2. 2
L-Data Plants
First Sergeant Leonid Asipov
Introduction 5
מדעית ואמינות מדעיות תוצאות הצגת
..............
5
Theory 6-15
צמחים של פוליטיקה ,פילסופיה
.................
6
הבוטניקה משימת
...................
6
7……………Botany T test
Two botany conceptions……………7
Enzymes : Structure which bonds substrates…………….8
Enzymes does not need any energy for
function…………….11
Energy = movement ……………….11
חום הולכת = ריאקטיביות
.................
12
האנזימים נבנים איך ,תבליטנות
..............
11
הפ החיידקים שלושת תאוריית
.וטוסנטיים
...............
14
צמחים אבקת
..........
15
חמניות תנועת
................
15
Criticism 17-75
Botany research is not needed anymore………..17
L-Data vs: Development of synchronized, autonomous,
and self-regulated oscillations in
plant……………………..18
No Oscillations……………………….32
Presentation Of CO2 Gas-exchange
experiments………..33
Plant screening systems……………33
3. 1
L-Data vs "Expression of cyanobacterial ictB in higher
plants enhanced photosynthesis and
growth"…………….34
LData vs "Enhanced photosynthesis and growth of
transgenic plants that express ictB, a gene involved in
HCO 3–accumulation in cyanobacteria"……………46
L-Data vs "A putative HCO transporter in the
cyanobacterium Synechococcus sp. strain PCC 7942
"……………….51
L-Data vs "CO2 Concentrating Mechanisms in Algae:
Mechanisms, Environmental"…………………55
Transgenic protein enhaces reactivity………………….56
L-Data vs "Changes in the properties of reaction center II
during the initial stages of photoinhibition as revealed by
thermoluminescence
measurements"………………………57
L-Data vs "Thermoluminescence and flash-induced
oxygen yield in herbicide resistant mutants of the D1
protein in Synechococcus PCC7942"………………58
The C4 plants photosynthesis is not existant……………59
Sekoia……………………………60
NOT N, possibly only water is reason for difference in
weight of wheat……….60
The Ci parameter is WRONG……………….…..60
L-Data vs “The Role of Tobacco Aquaporin1 in Improving
Water Use Efficiency, Hydraulic Conductivity, and Yield
Production Under Salt Stress1”..………….65
L-Data vs "SYNERGISM OF LIGHT AND FUSICOCCIN
ON STOMATAL OPENING" ………..……….74
Cactuses create sugar during the day………….……..75
Development 77-81
(Hebrew) Plant Apical Dominance Model………..77
4. 4
Plant Development Strategy………..80
There is no main growth control in plant : transpiration is
related to nutrient
transport………………………………….81
Stomata 83-96
Stomata : CO2 Triggers………………….83
Stoma aperture…………………92
CO2 and Stomata…………….93
Stomata do not affect photosynthesis, only
transpiration……….93
CO2 is trigger for stomata opening without any
biology………..95
Stomatal response to CO2……………..96
Water 97-105
The Role of the Difference Between Plasma and Vacuole
Membranes Water Conducting Properties : A Model
Approach…………………97
Plant Cappilarity…………………….101
Plant dehydration threshold with respect to transpiration
……………..103
The reason for "digital" effect of drought………………..105
Photosynthesis 105-107
Photosynthesis ……………..105
There is no CO2 fixation : O2 concentration blocks
photosyntesis by reverse CO2 formation ………………106
Stomata don't affect photosynthesis…………………..107
Patents 108-125
5. 5
Carbon Enrichment System………………..108
ייבו להגדלת צמחים של וויברציה
ל ……………….109
גשם מי לאגירת ……………………מערכת..110
אידוי בסיס על צמיחה לעצים ……………………רפסודה.110
Transpiration Measurement Method ……………..110
לעלים כלורופלסטים …………………הוספת.111
PLASTIC ABOVE THE FIELDS FOR WEED
CONTROL………………112
Water consumption measurement system
…………………112
בשקיות צמחים ……………………………גידול114
לצמחייה ………………………ביוב115
Greenhouse of the future………………….115
116
............
צימ
סוכר תוסף : בחושך צמחים וח
Growing plants in dark, industrial sugar production from
basic components CO2 and water…………………117
O2 growth enhancer…………………….118
Urine on leaves of plant to enhance rate of plant addition
of tissues……..119
Screening system……………….119
120
.........................
..
השקייה ניסוי
121
......................
לצמחים ביולוגי דשן
LIPID ON PLANTS TO CONSERVE
WATER………………..120
הזנת
צמחים
....................................
122
השקייה ללא : בים חממות
......................................
122
Grow on trees. Plants are well grown on tree sugars,
water and proteins…….123
Artificial plant…………………………………124
Transpiration measurement……………………124
מצמ חשמל רווח
ח
...............
............
125
7. 7
LEONID
11/19/2012 9:54:54 AM
קומוניסטים=ת צמחים
ו
אחר שדה תות ,אור ללא צמח אם גם שדה ת
סוכר שולח
ו
מים
.
צמח יש
גזע ערבוב בלא מאמנינים פשיסטים ים
.
מת
וגטטיביות ע"י רבים
.
גדול מרחקים :השאר=קפיטליזם
ים
של
בעקבות צימוח
צ עם ערבוב זרעים פיזור
מ
ויחידאיות מרוחקים חים
המשאבים מבחינת
.
הבוטניקה משימת
Leonid Asipov
12/19/2012 10:27:05 AM
הדל לכל שיספיק לסוכר צמחים צימוח
אידוי ,בים הצימוח .במדינה ק
חממות .השקייה מי חסכון ,בקרקע המים לעיבוי רשת דרך המים
גשמים בזמן מים איסוף ,השדה מעל
... (C)
אסיפוב לאוניד
Botany T test
Regular statics table of significance, is wrong for botany.
The table : arbitrary values of significance good for a
certain field of science. Other fields, other exceptions.
Botany has to create own significance table. Opinions of
all botanists has to be considered according to amount of
plants good for any results discussion and the amount of
plants enough for significance claims. The mathematic
values were arbitrary and new values for the statistics
significance table has to be proposed. The work of talking
with all
botanists is for a student.
Two botany conceptions
8. 8
Leonid Asipov
8/28/2011 12:56:11 PM
Two conceptions in botany and life sciences:
1 - Everything is controlled by biological
mechanisms.
Biological mechanisms presume biological molecules
acting as regulators on other biological molecules.
The biological molecules alter cell mechanics,
chemistry or alter particle construction.
2. There is no biological control. The visible control is due
to physical phenomenae such as diffusion rates.
The structure of the organism causes the visible control to
be. There is no actual control, and everything is diffusion
in effect of organism structure (such as leaf form, plant
height).
In some phenomenae both conceptions can explain the
basic principle. Reality can be to either of the theories.
Our goal is to develop both conceptions to a state of total
understanding.
The next stage is more experiments with real plants.
Enzymes : Structure which bonds substrates
Leonid Asipov
8/26/2011 9:40:05 AM
9. 9
Enzymes are the most important biological molecule.
The temperature or the speed of particles
of biological organisms cannot exceed little values.
The way to achieve reactions is holding of atoms
close to each other for a period of time.
During the time, a bond is created by the gravitational
force.
Other option is strong collision (happens not in bio
systems).
The way to hold the connection of two atoms/molecules is
to build a molecule with complementary shape with
option to bond partially and temporary.
This molecule is randomly bonding the substrates
and keeps them together for some time.
During this time the bond is created in lower
temperatures. The energy or movement of small
particles needed is for detachment from the enzyme.
The new bond is strong and the enzyme connection
and strength is low. The enzyme degrades during
short time. In the cell there is large amount of
enzymes of various substrates and this enhances
the possibility of bonds between substrates.
Any mechanical structure of enzymes is not likely.
The principle which stands micro levels is
complementary structure which randomly bonds
the substrates. No affinity is possible but the
complementary shape keeps the two
substrates together and bond is created.
11. 11
Enzymes does not need any energy for function.
Leonid Asipov
8/29/2011 6:22:13 PM
Enzymes does not need any energy for function.
Since speed is no longer needed for the activation
energy, no energy is needed. Activation energy
is only an energetic gap in the opposite direction.
If two molecules are held together, no speed is
needed and thus no energy.
The enzymes act as holding molecules.
The connection between substrates is made
by gravitation and enzyme degradation is
spontaneous since the molecule is unstable.
In order to disconnect the enzyme and the substrate,
small destructive molecules are needed.
Such molecules are called "reactivity".
Reactivity cause enzymes to degrade.
The degradation is needed for the detachment
of the enzyme and substrate.
Due to reactivity, biological molecules are temporary.
Energy = movement. Sugar=> small particles
enlarging heat flow. Energy needed for life=
movement or vibration.
17. 17
גרוויטציה = קפילריות
Vibration such as voice can accelerate diffusion. Particles
colliding with the vibration object can accelerate diffuse
quickly.
Flowers are located on ends of branches where
transpiration is large. Drawing of matter such as sugars,
proteins and fats are best on branches with large
transpiration. Transpiration draws water and matter
forward.
Criticism
Botany research is not needed anymore
19. 19
לאוניד אסיפוב
11/14/2012 6:27:04 PM
Leonid Asipov, CEO L-Data
P.S
I was a student of the publishers and the data abundant.
Please consider the papers I enclose scientifically.
SUMMARY
NO SHORT TERM OSCILLATIONS OF PLANT
TRASPIRATION EXISTENT. THE MEASUREMENT
EFFECT IS DUE TO MEASUREMENT INACCURACY
Flaws in the reported article.
1. SIMILAR OSCILLATIONS WITHOUT IMPORTANCE
DAY/DARK
2. NO SHORT TERM OSCILLATIONS AT GAS
EXCHANGE MEASUREMENT
3. OSCILLATIONS ARE SMALLER THAN THE NORMAL
INACUURACY
OF A ELECTRONIC SCALE: +- 2 grams (ACCORDING
TO PRODUCER)
Data from Tedea-Vishay 1040 C3 transducer. Total error :
0.02% of rated output. 0.02% of 5000 grams (average
plant) is +- 1 gram.
The total error of course is measured in ideal conditions of
humidity, noise, temperature and wind. Normal
20. 21
experimental conditions of course
Elevates the noise. THE NOISE OF WEIGHT SCALE IS
AT LEAST +-1 gram. Actual noise: +- 2 grams.
1. There is inability to differentiate between oscillations
due to fluctuation in light, temperature, humidity and water
availability and the reported "spontaneous" oscillations.
The reported experiments were held in a greenhouse with
fluctuating irradiation parameter. The claim for
"spontaneous" transpiration oscillations is impossible due
to unstable environment. The control is a wet blanket
positioned on a similar scale weight. The short term
fluctuations of the control is without any significant
correlation to plants, suggesting the short term fluctuations
are not related to temperature or irradiation, but another
sensor of transpiration (gas exchange of a single leaf)
suggests the total absence of oscillations on basis of short
term periods of less than 5 minutes. Since the
temperature or light was not measured by any other
sensor than the scale, an artifact can be speculated. The
control should have been additional sensor such as light,
temperature or humidity simultaneous to the weight
measurement. The absence of a proper control to the
experiment suggests improper scientific approach based
on a single device. The electronic scale has certain
accuracy. The minimal accuracy stands for 0.02 % from
the weight, and about +- 1 grams. The size of the reported
oscillations is about +- 1 gram. Similar to minimal
inaccuracy, suggesting the short term oscillations are
noise.
2. The weight measurement should have been held in
controlled environment, to avoid any fluctuations of light
as effect on transpiration. Since the experiment was held
21. 21
in unstable light, some of the oscillations may be due to
external irradiation fluctuations…
The experimental setup
Fig 1A: Whole plant on a weight scale for continuous
transpiration measurement. Readings every 10 seconds,
average to 3 minutes.
Fig 1B: Gas-exchange measurement of a single leaf.
Points are measured every 2 minutes.
22. 22
The size of the oscillations is similar to normal
measurement noise
Fig2: Weight scale measurement of a whole plant.
23. 21
Inaccuracies unseen since the total weight of the plant is
much larger than the noise.
Fig 3: Small noises are seen after differentiation since the
transpiration value between each point and the following
point is only ~10 grams. The noise is
0.02% of total plant weight (about +-1 grams). Noise is
20% of measured value. Significance of results exists
when value of transpiration is much larger than noise.
On scale of 30 minutes the extent of the noise is 2.5%.
After averaging on the range of 30 minutes, the
oscillations are correlated with the irradiation levels (Fig
6).
PAPER 1: DATA FROM MSC THESIS ACCEPTED BY
THE CAMPUS OF REHOVOT.
Simultaneous measurement of Gas-Exchange and
Scales. Additional sensor.
24. 24
The scale is only one sensor. Artifact regarding the scale
itself is to consider.
The control of the scale is transpiration measured by gas-
exchange method.
The oscillations are expected to be larger, since a single
leaf is less synchronized than the whole plant. The more
stomata is measured, the less oscillations are expected,
since the oscillations of single stomata are averaged.
The whole plant transpiration can be considered as NON
oscillating PARAMETER.
Experiment Setup
1. Load Cell A whole tomato plant is positioned on a load
cell in a water container. The rate of transpiration is
calculated from weight loss of the plant and the container.
2.Gas-exchange The rate of transpiration of the same
plant is simultaneously measured using gas exchange
method on a single leaf in the upper part of the plant. The
experiment was made on a well watered plant during a
whole day in natural light.
Results
25. 25
Fig 4: Simultaneous measurement of transpiration using
load cell and gas exchange methods.
Conclusions
The oscillations are the similar size during the night and
the day periods.
The forecast is that during high transpiration period, the
oscillations are supposed to be higher. However the
oscillations, are not different. The fact that the oscillations
are abundant during the night suggests that the
oscillations are due to measurement artifact, which is
basically inaccuracy of the weight scale.
The accuracy is 0.02 % which is about +- 2 grams.
The reported short term oscillations are about the same
size, ~+-1 gram.
26. 26
The lack of any oscillations during the night period
observed in gas-exchange measurement, suggests that
the oscillations are an artifact related to the electronic
scale.
During the day period, there are long-term light-correlated
oscillations seen also in gas-exchange experiment.
The fact that the experiment was held in natural light, adds
uncertainty to the experiment. There is a degree of
oscillations related to light fluctuations. The scale weight
oscillations are hardly differentiated from light oscillations,
however, the nocturnal oscillation of the weight suggests
that the oscillations are an artifact.
The better experiment is to measure in controlled
environment, with constant light levels.
The oscillations of a single leaf are expected to be larger
than the oscillations of a whole plant. Light-Driven
oscillations affect more higher leaves. The whole plant is
less affected due to shading of leaves. The following chart
consists of a smoothed (30 minutes moving average)
weight scale experiment and a simultaneous gas
exchange measurement.
Chart 1: Simultaneous measurement of plant transpiration:
Gas exchange –and scale.
From Maters thesis Leonid Asipov, 2011.
27. 27
Fig 5:
1: Light (ORANGE)
2: Transpiration of a leaf(BLUE)
3. Transpiration of whole plant(DARK BLUE)
Light: SUNLIGHT. LONG TERM FLACTUATIONS ARE
CLEAR IN GAS EXCHANGE AND SCALES.
Short term oscillations are random and night/day
unspecific. Longer oscillations (0.5 hour) are related to
fluctuations of irradiation levels. The pattern of the
correlation is seen both in gas-exchange and scales
experiment. Longer term oscillations are related to
irradiation fluctuations. The short term oscillations seen on
electronic scales measurement, are related to weight
measurement inaccuracy.
We would expect that oscillations in transpiration rate
28. 28
should be larger at daytime since transpiration is larger.
The measured oscillations were found to be similar during
DAY/NIGHT,
suggesting the oscillations are not related to the
transpiration rate but to spontaneous measurement noise.
SHORT TERM OSCILLATIONS ARE ONLY SEEN AT
ELECTRONIC SCALES MEASUREMENT OF A WHOLE
PLANT.
The significance of the measurement of the scales is
when the transpiration is larger significantly than the
measurement noise.
The transpiration of 0.5 hour is 80 grams at maximum
transpiration levels. The noise is 0.02 % which is about +-
1 grams. 2 grams is 1.25% of the measurement. The
smaller the scale of consideration, the degree of noise is
larger than the total transpiration during the period of time.
The transpiration during 3 minutes is 8 grams. 1 grams of
8 is 12.5%. The extent of the spontaneous oscillations is
large and the spontaneous noise on the scale of 3 minutes
(the considered time scale of the reported article) is similar
to the reported oscillations(~10% of the measurement).
Since the reported oscillations are similar to the
spontaneous noise of the electronic scale, short time
periods are not to be considered significant to the
measurement. The measurement is considered to be
significant when the extent of the transpiration is
significantly larger than the noise. The time scale to
consider is larger than 0.5 hour.
The short term fluctuations are artifact.
29. 29
Fig 6:
Measurements each 10 sec, averaged to 3 minutes.
Y Axis: weight loss [grams/sec]
X Axis: Time of day
PURPLE: Transpiration without smoothing
GREEN: Smoothing 13 points
RED: Smoothing 25 points
"The term" short term oscillations are related to
fluctuations of transpiration on scale of less than 3
minutes. The short term oscillations are considered an
artifact of the electronic scale.
Transpiration and Photosynthesis measurement in
controlled environment: stable transpiration parameter,
changes due to CO2 changes in the air.
GAS EXCHANGE METHOD SHOW NO SHORT TERM
30. 11
OSCILLATIONS IN CONTROLLED ENVIRONMENT
1. Gas-exchange measurement of photosynthesis and
transpiration in controlled environment: no short term
oscillations, transpiration responds to changes in CO2.
Fig 7:
1: orange Light levels.
2. Purple CO2 Concentration
3. Green Photosynthesis
4. Blue Transpiration of a tomato plant is controlled room.
Measured by gas-exchange method.
31. 11
Fig 8:
Gas-exchange measurement of photosynthesis and
transpiration in controlled environment: no short term
oscillations, transpiration responds to changes in CO2.
Similar experiment
Gas exchange method works on a single leaf. The
speculated oscillations should have been larger since
synchronization of oscillations is hard to imagine.
Nevertheless, no short term oscillations in plant
transpiration are evident in gas-exchange measurement of
transpiration.
CONCLUSION: THERE ARE NO SHORT TERM
OSCILLATIONS IN PLANT TRANSPIRATION
L-Data http://www.ldata.co.il
33. 11
Presentation Of CO2 Gas-exchange experiments
Leonid Ldata
10/10/2011 9:43:59 AM
Proper presentation of CO2 gas exchange measurements
has to be CO2R-Photo and CO2R-Transpiration.
Always on the x axis, a variated parameter has
to be presented.
All comparable points has to share one X value.
On a single x value Y values represent different
experiments. Plants are compared by photosynthesis
and transpiration at same CO2R (external CO2
concentration).
Two charts are to be presented: One for
Transpiration and one for photosynthesis.
Conclusions can be made on both physiological
parameters while each is presented as function
of CO2 in the air.
Plant screening systems
Leonid Asipov
11/15/2012 12:37:42 PM
Plant screening systems
The bad systems
1. Gas exchange
Limitations : Small count of measurements are possible.
34. 14
Only one measurement at one time. Different
measurement daytime. Different age of plants. Unnatural
light usually the method. The cold artificial light is not
transpiration inductive as sunlight.
MASS PLANT SCREEN IMPOSSIBLE. Result : studies
based on few plants and insignificant results.
2. Weight scale measurement. Due to humidity, the weight
of the plants rises. The rise in unpredictable due to
instability of temperature, irradiation, and humidity.
The good system
Real water volume measurement.
Dry weight measurement.
WUE= DRY WEIGHT[g] / TOTAL WATER[g]
Larger transpiration, LESS WUE.
WITH LESS WUE BUT LARGER MASS OF PLANT,
better grow more ORGANISMS WITH HIGH WUE.
L-Data vs "Expression of cyanobacterial ictB in higher
plants enhanced photosynthesis and growth"
Leonid Asipov
11/15/2012 10:57:26 AM
From letter to the editorial director of the magazine…
"My Name is Leonid Asipov. I am the co-author of the
35. 15
article and the worker which accomplished the
experiments. The data results are insignificant and no
conclusions are possible on basis of the results. Whole
experiments were neglected by the main author of the
article and the conclusions are scientific lies… "
TOPICS
1. ONLY PARTIAL DATA WAS PRESENTED. THE
ADDITIONAL PLANTS SHOW NO SIGNIFICANCE
RELATIVE TO WT.
2. SECOND GENERATION OF PLANTS, GROWN OUT
OF THE BEST 1-ST GENERATION PLANTS, SHOW NO
SIGNIFICANCE AT ALL.
3. FALSE CONCLUSION IN ABSTRACT OF THE
ARTICLE, ACCORING TO THE AUTHOR,
THE ENHANCEMENT OF PHOTOSYNTHESIS IN ONLY
AT LIMITING CO2 CONDITIONS,
HOWEVER, THE RESULTS (BASED ON ONLY 1
PLANT) SHOW THAT AT ALL CO2 LEVELS, THE
TRANSGENIC PLANTS ARE BETTER. NOTE THAT
MOST OF THE DATA (THE REST 36 TRANSGENIC
PLANTS) WERE NOT PUBLISHED.
4. No significance tests were calculated. The group of
transgenic plants is too small(4 plants).
THE REAL DATA: EMPIRIC MEASUREMENTS OF GAS-
EXCHANGE OF LEAVES: WT VS TRANSGENIC
PLANTS.
Time of day during measurement
36. 16
The measurement is Gas-Exchange of single leaves.
Transgenic and wt plants are similarly measured by the
gas-exchange device, in elevating CO2 concentrations
and in similar temperature and light conditions. The time
of day of the measurement of each plant was different.
Part of the plants were measured during morning and part,
during the evening.
The difference of the Photosynthesis rate between the
different times of day is great. During the evening, the
plants were photosynthesizing many hours already and
part of the enzymes needs regeneration. Moring is hours
of highest photosynthesis rates. Note that there is large
differences between the photosynthesis and transpiration
rates of the plants. Part of the large differences is due to
different time of day during measurement.
Scientific conclusions
The plants are to be measured in equal conditions: equal
time of day and equal age. When the experiment is large,
the task is impossible. Since the gas-exchange
measurement device is based on a single leaf and is
expensive, in addition to long measurement time of the
CO2 curve (at least 15 minutes for each plants +
acclimation time) and large number of plants demanding
other method for plant screening for better photosynthesis
and transpiration properties. Measurement in a single
CO2 concentration is partial solution for possible
measurement of many plants in short period of time.
The results of the article: partial data was presented.
All the transgenic plants are to be considered similar
"treatment" and a large number of plants are to be
measured. T significance tests are to be calculated. If
some particular transgenic plant is for any reason is
37. 17
considered to be "other treatment", the plant is to be
cloned to next generation and large amount of plants are
to be measured in order to perform T significance tests.
Second generation of arabidopsis(NOT PUBLISHED
DATA)
Second generation was cloned of the 4 top considered
transgenic plants and the results showed no significant
difference relative to wt plants.
THE PAPER WAS PUBLISHED WITH WRONG
CONCLUSIONS BASED ON PARTIAL DATA, WHEN ALL
DATA WAS ABUNDANT. THE SCIENTISTS ARE LIERS.
Presentation of all the data
The real Gas-Exchange data. The first experiment
included 37 transgenic arabidopsis plants vs 7 wt plants.
The published chart however consists of only 1 transgenic
plant. The researchers have neglected all the rest 36
plants and CONCLUDED FALSE conclusions, according
to which the transgenic protein has positive effect on
photosynthesis of plants.
The absence of statistics
The variation of the plants is high and T significance tests
of all the 37 plants vs wt show no significance at all. The
experiment of protein transformation is to be considered
similar “treatment”. The 4 top plants were cloned to the
next generation and similar tests were conducted. No
significance was seen.
Original chart published in a seminar: only 4 plants of 37
transgenic plants were published.
38. 18
In the paper, only the top plant (1) was published. In the
article Y standard deviation bars of wt did not appear.
Note the inconsistent conclusion in abstract of the article :
“photosynthesis was enhanced under limiting and not
under saturating CO2 concentrations…” The first
(published) transgenic plant is better photosynthetically at
ALL CO2 points(NOTE THE REST 36 PLANTS WHICH
ARE NOT).
All 37 transgenic plants. Only top 1 was published. The
rest of the plants are with large variance. Better or worse
than wt. Wt is also variant. Note that only 7 wt plants were
measured. More plants, more variance.
Photosynthetically, Only the top (1) plant is better then wt.
39. 19
THE PUBLISHER DID PUBLISH ONLY 1 TRANSGENIC
PLANT. REST OF THE DATA WAS HIDDEN. THE
SCEITIFIC FRAUD WAS CAUGHT!!!!!!!!!!!!!!
2nd generation arabidopsis (number of plants > 30).
UNSIGNIFICANT DIFFERENCES BETWEEN WT AND
THE TRANSGENIC.
40. 41
The protein was also tested in other plants. Similar gas-
exchange experiments were conducted.
The protein does NOT enhance photosynthesis or prevent
transpiration…
Soybean experiment.
The theory behind the article is wrong.
41. 41
Theoretic criticism.
The CO2 channel in chloroplast is not needed : the
membrane keeps the unique environment in chloroplast.
Channels conduct to and from the chloroplast. The reason
of the membrane is to block particles from diffusion from
the chloroplast to the cytoplasm.
Additional CO2 channel in chloroplast membrane would
have forced diffusion of small and medium particles from
the chloroplast to the cytoplasm, slowing the enzymatic
processes in the chloroplast.
The diffusion of CO2 to chloroplast is fast due to large
surface area/volume ratio. The main slowing process is
diffusion through the cytoplasm.
The surface area of a chloroplast is large relative to
volume…. The main obstruction to diffusion of CO2 from
the air the chloroplast is cytoplasm volume and cytoplasm
membrane… The distance of diffusion is the whole
cytoplasm… The distance of diffusion through the
chloroplast is small and don't forget the large surface area
/ volume ratio of the chloroplast which cause fast diffusion
through the membrane. The conclusion is that no CO2
channels are needed to chloroplast membrane. Addition of
CO2 channel, don't enhance any processes in the plant
cell…And might slow some processes by diffusion of
needed particles from chloroplast to the cell cytoplasm.
Bacterial proteins does not express properly in higher
organisms. Note folding proteins which are needed.
Expression of bacterial protein in higher plants: wrong
concept. It is well known in science that proteins from
42. 42
eukaryotes cannot be easily expressed in prokaryotes. In
order to create the antibodies, mammal or fly biological
systems are needed. The 3rd structure of the proteins
need folding proteins. Spontaneous expression of a
bacterial protein in plants is not likely to produce a proper
protein. The folding proteins of the plant are not
compatible. A whole set of additional proteins are needed
in order to create a proper protein. The ribosomes of
bacteria are different, suggesting the building of the
proteins is as well. Possible even the genetic code differs
and codons code to other amino acids.
The task of the researchers is to prove the protein
function, 3-d structure and proper attachment of the
protein in the chloroplast membrane.
Since the protein was described as "putative", and no 3d
structure was proved, as well as no positioning in the
chloroplast membrane, it is impossible to claim any
biological effect. The foreign bacterial protein may be
malfunctional and not even expressed in chloroplast.
The difference of the plants in sampling time
The experiment was sampled during 2 months and life of
arabidopsis plant is about 3 months only. The first plants
were the best transgenics, the control plants were
measured during the last 4 days of experiment. The
control plants were 2 months older than the best
(published) transgenics. The difference of the plants is
mostly at transpiration parameter, not photosynthesis, and
the root system of the transgenic plants which are older by
two months is obviously larger, and thus the transpiration.
The difference between the transgenic and wt plants is
due to age, no genetic differences.
External CO2 vs Photosynthesis (wt vs best transgenics)
43. 41
External CO2 vs transpiration: transpiration of wt plants is
larger (most possibly because of root system)
44. 44
The ages and sampling dates :
PROPER SCIENTIFIC WAY
The proper way of molecular biology is protein
crystallization to prove proper function : CO2 channel.
CO2 channels, as any channel is a pipe-like protein. The
pipe is only molecule size specific. CO2 channels conduct
medium and small size molecules such as water, CO2, H
and other.
Proof of protein expression and positioning in chloroplast
membranes. The protein has to be proven to have proper
channel function and positioning.
45. 45
ONLY AFTER PROPER MOLECULAR PROOF,
transgenic plants are to be grown and compared with wt
plants. BEFOREHAND we cannot even ask any scientific
question, since the protein may be not a channel, not
properly built by the plant protein building machinery or
not to be positioned in the chloroplast membrane.
GUESSING is not proper science.
THEORY HAS TO PERCEDE the scientific research with
all the possibilities of the results.
The Ci parameter : wrong science
The Ci parameter is wrong. Photosynthesis is linked to X
and Y axis on Ci-Photo charts.
Proper comparison is possible between curves is possible
at similar X points. The empirically changed parameter is
the external CO2 concentration (CO2R). Photosynthesis
and Transpiration are biological parameters affected by
the external CO2 change. The proper way to compare the
effect of external CO2 on photosynthesis and transpiration
is to present the external [CO2] on the X axis and plot 2
separate curves for transpiration and photosynthesis.
The Ci parameter is wrong :
Ci =
External [CO2] * STOMATA APERTURE AS
TRANSPIRATION * pressure /
PHOTOSYNTHESIS * leaf temperature
One Ci, different Photosynthesis and transpiration values.
If transpiration and photosynthesis is high, the ci is same if
transpiration and photosynthesis is low.
46. 46
When plotting relative to photosynthesis, points are
compared at different X values, which are related to Y
(PHOTOSYNTHESIS) .
The ci parameter cannot tell anything about the real
properties.
Since it is basically ratio of
TRANSPIRATION/PHOTOSYNTHESIS.
PHOTOSYNTHEIS and TRANSPIRATION IS TO BE
PRESENTED AS FUNCTION OF EXTERNAL CO2
CONCENTRATION.
LData vs "Enhanced photosynthesis and growth of
transgenic plants that express ictB, a gene involved in
HCO 3–accumulation in cyanobacteria"
Leonid Asipov
11/15/2012 11:46:40 AM
Under low relative humidity, there is more CO2 abundant
and therefore the stomata are more closed.
Photosynthesis is enhanced.
Under high humidity concentrations, there is less
concentration of CO2 in the air, and thus photosynthesis is
declined.
47. 47
We would expect the protein to enhance at higher CO2
concentrations,
When CO2 is a limitation.
The results are opposite to the expectation. Our
conclusion that the article is a
scientific error.
Internal article inconsistency.
Since there almost no PHOTOSYNTHESIS dependence
on stomata aperture
(Fig 1),
The photosynthesis under any humidity is relatively
constant. The only factor is humidity which lowers the CO2
concentration of the air. According to the article, at lower
CO2 concentrations, WT plants are better. WT is the
better CO2 conductor than the transgenic plants.
The "faster" growth of the transgenic plants at lower
humidity, most possibly is accompanied with higher
transpiration rates. Why the Water USE Efficiency
parameter was not calculated?
The transpiration was reported to be similar between the
wt and the transgenic plants. Why more growth at lower
humidity, at higher CO2?
"CO2 compensation point": the plant is breathing more
and thus there is more CO2.
More energetically demanding plant.
The activation point is lower at transgenic plants, proving
48. 48
more cell breathing, thus more CO2 near the chloroplasts
and thus earlier photosynthesis activation.
The real DATA
The number of plants is only 4 transgenic plants vs 1 wt.
How is it possible to conclude anything of so few plants?
Part of the transgenic plants are not better, how to
explain? If part of transgenic plants are to consider other
properties than the rest of the plants, the plants should
have been cloned to the next generation and large
number of plants were supposed to be sampled.
Conclusions are to be made on basis of T significance
tests of many plants and not on only 4 transgenic and 1 wt
plant. The Arabidopsis conclusion was based on 2
transgenic and 1 wt plant. Conclusions could not have
been made due to too small plants group. The wt plants
are relatively at the average of the transgenic plants CO2
curve. Why the conclusion was that the transgenic plants
are better, when some transgenic plants were not?
Why DO we care about photosynthesis at limiting
conditions. The real conditions at fields are not limiting.
There is no advantage of transgenic plants and NO
transpiration difference.
The stomata respond to CO2 with closure. Better
photosynthesis, lower CO2, more stomata aperture and
MORE TRANSPIRATION. If the plants are better
photosynthethicly at lower humidity, MORE
TRANSPIRATION is EXPECTED but was not reported.
CO2 is the main trigger of stomata aperture, and
photosynthesis is related to CO2 concentration in the leaf
49. 49
and near the stomata. At high photosynthesis, more
transpiration is expected.
DATA INSTABILITY:
There was too many samples and the AIR CO2
concentration at every sample is not clear. Either the
researchers were only slightly changing the AIR CO2
concentration, or there was no enough time of acclimation
to the external CO2 conditions. The differences in Ci is at
almost every sample, was there a real change is the air
CO2? MORE ACCLIMATION TIME IS TO BE GIVEN TO
THE PLANT AFTER CO2 changes in the air. The Ci
parameter has to be constant and only then the sample to
be taken. There are more than 20 samples of every plant
and the concentration of CO2 was changed between 50
and ~800 (UNMENTIONED AT ARTICLE) . Was the
external CO2 changed at every point? Why there is more
than one sampling at a single CO2 point?
Acclimation times and CO2 concentration points were
unmentioned at Materials and Methods.
Growth results
There was too few plants to conclude any conclusion.
Conclusions are to be based on large number of plants
and statistical T tests are to be performed.
The growth in lower humidity (which is seen as higher
photosynthesis measured at the external to leaf air) would
have affected transpiration. Higher CO2 of the air is
causing stomata aperture to grow, and thus the
transpiration rate is HIGHER.
The transgenic plants are more water DEMANDING,
which is not positive to agriculture.
50. 51
"The experiments were held 6 times", where are the
results? Why the statistical data was not presented?
70% RH is wide and common humidity. Under many
agricultural applications there is no advantage of the
protein. WHY TO ADVANCE PARTIAL AND UNCLEAR
SOLUTION which is not better at most cases?
Conclusions
1. The better photosynthesis at lower humidity is not
logical. There is more CO2 in the air. Better CO2
assimilation is of WT plants. The protein enhances CO2
transport only at higher CO2 concentrations.
INCONSISTENCY. The transgenic plants do not grow
better at lower co2 concentrations, but at higher ones.
2. The limiting CO2 concentrations are not real at earth.
Normal air has no limitation of CO2 (~400ppm). No
advantage of the protein.
3. NO EFFECT ON TRANSPIRATION WAS REPORTED,
which is impossible. More photosynthesis, more stomata
aperture, since stomata are triggered by CO2
concentration.
4. Higher transpiration rates are expected from transgenic
plants due to higher photosynthesis. The transgenic plants
are WATER DEMANDING.
5. The water use efficiency was NOT CALCULATED.
6. Experiments were based on SMALL NUMBER OF
PLANTS.
7. NO STATISTICS WERE PRESENTED.
51. 51
CONCLUSIONS ARE IMPOSSIBLE BASED ON ONLY 4
TRANSGENIC AND 1 WT PLANT.
TRANSGENIC PLANTS AS PRESENTED ARE MORE
WATER DEMANDING AND GROW BETTER AT HIGHER
CO2 CONCENTRATIONS (LOW HUMIDITY)
L-Data vs "A putative HCO transporter in the
cyanobacterium Synechococcus sp. strain PCC 7942 "
Leonid Asipov
11/15/2012 11:54:34 AM
CO2 CHANNELS ARE NOT NEEDED IN BACTERIA
Bacteria have large surface area/volume ratio, and thus
there is no channels needed to conduct water or CO2.
The mutant, which needs more CO2 for proper
photosynthesis rate, can be defected in any
photosynthetic-related protein, not especially CO2
channels.
If there are CO2 channels in bacteria, why to add a
channel to a chloroplast?
The channels in bacteria would have forced small
molecules such as H, H2O, CO2 and many more
molecules to diffuse out of the bacterium. The cytoplasm
wants to remain UNIQUE and thus any channel force
changes in needed particle concentrations.
EUKARYOTIC cells are large, with SMALLER surface
area/ volume ratio, and thus channels are necessary.
A CHLOROPLAST IS A CYANOBACTERIA.
CHANNELS ARE CONSISTENT.
52. 52
THE TERM "CCM"
"CCM" CO2 concentrating mechanisms. It is impossible to
concentrate CO2, ONLY to transport faster to the cell. The
diffusion rate is related to concentration of CO2 and to the
speed of conduction of CO2 by the cell wall. There is no
artificial concentrating mechanisms, only diffusion.
THE PROTEIN MIGHT BE NOT A CHANNEL
Since the "putative" protein is not necessarily a CO2
channel, which are not expected to be expressed in
bacteria, expression in plant will not initiate any change.
Plants don’t express the needed folding proteins for
bacterial protein and thus the protein might be not
functional.
First the structure of the protein to BE crystallized and
proved AS a CHANNEL.
THEN, the protein has to be proved to be positioned in the
chloroplast membrane.
ERROR IN FIGURE : "External Ci" concentration. External
= CO2R. There is NO Ci in cyanobacteria. Ci is
concentration of CO2 in the leaf.
Theoretically, the transporter wouldn't have influenced the
concentration, only rate of acclimation to the CO2 change
surrounding the cell. The external concentration of CO2 is
large (mmol) and the photosynthesis is slow (micromoles)
and therefore, after change in external CO2, there is
acclimation during which the inner cell concentration
becomes as the external concentration of CO2. The
53. 51
channel affects the length of the acclimation, during which,
there would have been differences in photosynthesis rates
between the acclimated and non acclimated bacteria.
After acclimation, the rate of photosynthesis is alike.If the
protein is not a channel, differences are expected even
after the acclimation.
The uptake is slow, and during uptake, the cell
concentration Barely changes. The rate of diffusion of co2
is LARGE, therefore, the cell concentration is CONSTANT
and similar to OUTER.
If the acclimation time after change in external CO2 is
long, there should be no difference between the mutants
and the wt. Since in the mutant there is already full steady
state photosynthesis (starting at 15mM HCO3 or 6 mM
CO2), the rate of diffusion is BEYOND the scope of the
limitation of photosynthesis. The real limiting factor of the
mutant relative to wt is SOME PROTEIN NEEDED FOR
PHOTOSYNTHETIC REACTIONS. The diffusion of CO2
is already at saturation, and all the cytoplasm is with
concentration of CO2 similar to the external. Since, the
mutants need more CO2 and not only more acclimation,
the mutants are impaired in photosynthesis Proteins
expression. LESS PROTEINS, LESS
PHOTOSYNTHESIS.
HIGHER CO2 FOR SILIMAR PHOTOSYNTHESIS.
TIME BETWEEN THE MEASUREMENTS(CHART 2)
WHAT WAS THE TIME BETWEEN
MEASUREMENTS(AFTER ALTERATIONS IN
EXTERNAL CO2)?
In CASE of the protein as a CO2 channel, more
54. 54
acclimation, would result in similar PHOTOSYNTHESIS at
similar [CO2]. At shorter times, there is still uneven [CO2]
between the surroundings and the cytoplasm. More CO2
needed for faster diffusion and similar Photosynthesis. In
case, time does not force any similarity of
PHOTOSYNTHESIS of wt and the treatment, THE
PROTEIN IS NOT A CHANNEL .
"The protein encoded by ORF467 contains 10 putative
transmembrane regions and is inner-membrane-located."
WHERE IS THE PROOF?
A PUTATIVE REGIONS WHICH ARE SIMILAR TO
OTHER STUDIED PROTEINS ARE NOT TO BE
CONSIDERED AS PROOF FOR PROTEIN FUNCTION.
THE DATABASE ANALYSIS OF PROTEINS ARE
MERELY A CLUE TO THE REAL PROTEIN FUNCTION
AND NO CONCLUSIONS CAN BE CONCLUDED FROM
ANY FORMAL SIMILARITY OF THE GENETIC CODE
AND ANY KNOWN PROTEINS. THE MOTIFS ARE NOT
PROOF BUT SPECULATIONS.
REAL PROOF = CRYSTALLIZATION AND POSITIONING
OF GFP FOLLOWING MICROSCOPY.
Channels/ carriers
NO ACTIVE CO2 CHANNELS ARE EXPECTED. THE
OUTER CO2 is mmoles, and the photosynthesis is
micromoles. THEREFORE, the DIFFUSION IS NOT
ACTIVE.
The "channels" are pipes which enhance membrane
transport. The pipes are unspecific.
The size of molecule is the only limiting factor. Large
55. 55
Surface area/ volume of bacteria,
lead to the unnecessary existence of any channels in
bacteria.
CONCLUSIONS
1. The protein might be not a channel.
2. If there are channels in bacteria, there are also
channels is chloroplast.
3. NO PROOF FOR FUNCTION
4. NO "CONCENTRATING MECHANISMS" EXIST
5. ACCLIMATION BETWEEN MEASUREMENTS
UNMENTIONED.
6. NO PROOF FOR POSITIONING
L-Data vs "CO2 Concentrating Mechanisms in Algae:
Mechanisms, Environmental"
Leonid Asipov
CCM mechanisms are not likely. CO2 is high
concentration in the air, relative to plant. Artificially to
elevate concentration is an impossible mission with an
inefficient nature.
Why to artificially elevate the concentration of CO2 in the
chloroplast? The chloroplast's nature sugar consumption
produces more CO2, thus elevating the CO2
concentration near the chloroplast. The discussed CO2
may be the reason scientists thought that the chloroplast
possesses CO2 concentrating mechanisms.
56. 56
The nature works with diffusion. It is impossible to
artificially change concentrations opposite to the
substance concentrations. Water, salt, CO2, sugar, all
substances diffuse naturally in the plant and the plant
adapts to certain concentration of the substances.
The citated work does not prove existence of CCM, but
states the CCM may exist. The science does not like
science fiction.
CO2 channeling. Channels are proteins which conduct
particles according to the concentration gradient. The
PIPES, are normal protein pipes.
There are no transporters but PASSIVE, with GRADIENT.
ENERGY IS MOVEMENT. MATTER DISSOLVING TO
SMALLER PARTICLES ENLARGES HEAT
CONDUCTANCE. THE DIFFUSION OF ONE
SUBSTANCE IS BARELY RELATED TO THE OTHER.
THE DIFFUSION IS RANDOM. THE DEPENDENCE IS
ONLY AT HIGH CONCENTRATIONS, WHEN THE
VOLUME IS A LIMITING FACTOR. THUS, NO ACTIVE
TRANSPORTERS EXIST. REGULAR DIFFUSION ONLY.
THE PLANT IS BUILDING THE ORAGNIZM WITH
PARTICLES SUCH AS SUGAR, PROTEIN OR FAT, NO
"ENERGY" EXIST IN MATTER. SOME REACTIVITY
(HIGH HEAT CONDUCTANCE DUE TO SMALL
PARTICLES) IS NEEDED FOR DISSIPATION OF
SUBSTRATE FROM ENZYME OR TEMPORARY LIFE
OF BIO SUBSTANCES, WHICH IS VITAL FOR NEW
CELL REBUILDING.
Transgenic protein enhances reactivity
57. 57
Leonid Asipov
30/12/2012 13:44:30
L-Data vs protein.
The expressed transgenic protein elevates reactivity which
is larger amount of smaller molecules causing elevation of
heat conduction, thus temperature on sunlight. More
transpiration and photosynthesis due to higher
temperature only and also more breathing resulting more
CO2 by product. This explains larger photosynthesis at
smaller CO2 concentrations of the air.
L-Data vs "Changes in the properties of reaction
center II during the initial stages of photoinhibition as
revealed by thermoluminescence measurements"
Leonid Asipov
11/15/2012 11:56:53 AM
Leaves of plants exposed to intense light accumulate
more light particles. At other temperatures, the particles
will be depleted off the plant tissue. The switch of the band
from higher temperature to 15% is due to more light
particles accumulated in the leaf.
Phosphorus is the main matter accumulating light and
producing phosphorylation. The concentration of
phosphorus is the main factor of phosphorylation, as
measured by TL system. The TL is mainly a phosphorus
meter. Additional light absorbing substances are also
existent.
A regular spectrophotometer can usually replace TL.
58. 58
Photo inhibition is due to destruction of plant leaves by
excessive irradiation. The cure: shading.
L-Data vs "Thermoluminescence and flash-induced
oxygen yield in herbicide resistant mutants of the D1
protein in Synechococcus PCC7942"
Leonid Asipov
11/15/2012 11:59:01 AM
L-Data vs
Thermoluminescence and flash-induced oxygen yield in
herbicide resistant mutants of the D1 protein in
Synechococcus PCC7942
Leonid Asipov 25.10.2012
Mutants which are different in light absorption, will reveal a
change in TL results.
More LIGHT PARTICLES ABSORBED, less temperature
of RELEASE.
NEUTRAL TO LIGHT MUTANTS will lead to SIMILAR TL
PATTERNS.
PHOSPHORUS ABUDNANCE = MAIN STATIC REASON
FOR FOLURUSCENCE AND ILLUMINESENCE OF
PLANTS IN DARK.
59. 59
The C4 plants photosynthesis is not existant.
Leonid Asipov
11/15/2012 12:07:02 PM
The C4 plants photosynthesis is not existent.
ALL PLANTS ARE "C3", usual photosynthesis of a
chloroplast. Chloroplasts are the photosynthetic bacteria.
The bacteria are independent, and is not counting on
bacteria in other plant cells.
The theory of C4 plants is not consistent with the large
rate of photosynthesis of C4 plants.
If the "Dark" reactions and the "light" reactions are at
different cells, which are at some distance, the diffusion of
the "light" reaction products: C and H, would have slowed
the photosynthesis by seriously. The C and H are results
of light effect on CO2 and water. At short period of time,
AN ENZYME has to bond the resulting molecules and
create sugar. Otherwise, the C and H recreate CO2 and
Water. Diffusion to other cells is not practical due to the
needed large concentration for the diffusion. There are
many types of cells and the diffusion is not especially
conducted to the specific "dark" reaction cells. Rebinding
of CO2 and water is likely, and therefore the rate of
photosynthesis is expected to DECLINE.
The explanation of the insensitivity to O2 of C4 plants is
cell structure. Bigger cells with larger stomata and more
cell breathing, which is O2 demanding. The "c4" plants,
Are more energetically demanding, sugar demand is high,
thus breathing, and thus less O2 is abundant. A strategy
60. 61
of plant which is rich in sugar.
Sekoia
wep Leonid Ldata
11/29/2011 4:23:11 PM
= דפוזיה מלבד עילוי כח שום אין כי עולים המים : גדולים עצים
הריכוזים להבדלי בהתאם חלקיקים של אקראית תנועה
.
יותר גדול באוויר המים ריכוז כן אם אלא ,ללחות חשיבות אין
.
.בעלים חורים מלבד מים של להובלה מערכת שום אין
NOT N, possibly only water is reason for difference in
weight of wheat
Leonid ASIPOV
10/10/2012 3:00:42 PM
"Near pipes there is more water and N. Weight of wheat is
higher. The authors wrote N is causing the weight. WHY
NOT IRRIGATION?
The Ci parameter is WRONG
Leonid Asipov
11/15/2012 12:21:54 PM
Mathematical function is set of points to compare at Y axis
at similar X.
The Ci parameter includes the photosynthesis.
WHY TO INCLUDE TO X THE Y axis of the CI CURVE?
61. 61
The Ci curve is comparison of PHOTOSYNTHESIS
VALUES AT DIFFEARENT Ci,
When Ci includes already photosynthesis, resulting
change in X values to different values.
The X of the compared points is not same. COMPARISON
IS WRONG.
The calculation of Ci is artificial (not measured) calculation
of a parameter similar to the measured parameter CO2R
(External CO2). The true parameter which should be set
to X axis is CO2R. CO2R is the manually or automatically
changed parameter and as function of CO2R changes in
photosynthesis and transpiration do happen in the plant
leaf.
The artificial parameter Ci, is a value close to CO2R, and
based on Transpiration/ photosynthesis ratio, leaf
temperature, and pressure. The artificial CO2
concentration is similar to CO2R, but less usually. The
ERROR of Ci is that the CO2 concentration of the leaf is
not related to STOMATA APERTURE. The diffusion of
CO2 is barely limited by stomata. The only effect of
stomata is on transpiration and Ci includes transpiration
as function of the CO2 concentration in the leaf.
Therefore, the Ci is artificial and wrong. See
"DECOUPLING OF PHOTOSYNTHESIS AND
TRANSPIRATION AFTER ALTERATIONS IN EXTERNAL
CO2".
62. 62
ORANGE: IRRADIATION
PURPLE: CO2 CONCENTRATION OF AIR
GREEN: PHOTOSYNTHESIS
BLUE: TRANSPIRATION
Figure 1: DECOUPLING OF PHOTOSYNTHESIS AND
TRANSPIRATION AFTER CHANGES IN CO2(Chart from
Leonid Asipov MSc THESIS).
The transpiration changes during a long time of 30
minutes or more after the change in CO2.
Photosynthesis changes almost instantly(30 seconds),
During the change in the transpiration rate, the
photosynthesis does not change. During the period of
change of the transpiration, Stomata change the aperture,
process which does not affect the photosynthesis rate.
The conclusion is that stomata mainly affect the
transpiration rate due to differences in concentration
63. 61
between the leaf the air and slower speed of water
diffusion (MORE LIQUID THAN CO2). CO2 diffusion is
fast and almost not limited by stomata.
The BEST parameter is the real parameter CO2R,
presented with Transpiration or Photosynthesis on
separate curves.
Ci unclear results : no conclusions
EXTERNAL CO2 VS PHOTO
64. 64
EXTERNAL CO2 VS TRANSPIRATION
CONCLUSION : TRANSIRATION IS THE CAUSE FOR
CHANGE OF CI , NOT PHOTO.
65. 65
L-Data vs “The Role of Tobacco Aquaporin1 in
Improving Water Use Efficiency, Hydraulic
Conductivity, and Yield Production Under Salt
Stress1”
Leonid Asipov
11/15/2012 11:34:24 AM
Rationale:
Expression of an additional aquaporin improves water
transport of the plant. Improvement of the water transport
elevates the transpiration levels. The photosynthesis
levels might elevate a little, however the main effect of
stomata are on transpiration.
Fig 1:
66. 66
1: light levels
2. CO2 concentration
3. Photosynthesis
4. Transpiration
Gas exchange experiment in controlled environment.
Tomato photosynthesis and
Transpiration rates during changing CO2 concentrations.
Result
1. Fast acclimation of photosynthesis to CO2 alterations
and constant photosynthesis rate during the stomata
aperture change.
Conclusion
1. Stomata barely affect photosynthesis.
2. The negative effect of salt on photosynthesis is not
result of stomata closure (which does not change the
abundant CO2 concentration), but due to the direct effect
of salt on the plant photosynthesizing enzymes.
3. Enhanced water transport may improve enzymatic rates
ONLY at salt stress conditions. The more open stomata is
due to the expressed aquaporin, and the result is
enhanced transpiration. There is no more photosynthesis
under normal salt conditions and thus, the stomata
aperture is not relevant to the reason of why under salt
stress there is a difference.
67. 67
Salt stress is effect of salt on plant enzymes in a way that
photosynthesis declines. Better water transport is a
solution to the problem. However, the high transpiration
rates causes the solution to be too water demanding. Salt
accumulation is another argument against the transgenic
plants. The water use efficiency declines and the protein is
not for commercialization.
The reported WUE enhancement of the transgenic plants
is absurd,
Water use efficiency: Total dry plant to unit of water
weight.
Plant dry weight [kg] /WATER [kg]
The transpiration rate is about 70 times faster (per mole
water relative to mole CO2) than photosynthesis.
In order to preserve constant WUE at elevation of 15% in
transpiration levels, which is seen in the transgenic plants,
photosynthesis has to be elevated 10 times (1000%) 70 +
15% of 70 = 80 photosynthesis units.
The molar weight of water is 19 and the assimilation of a
single C atom is only 6 gram/mole. The O2 is returned
back to the air and thus, at single molar measurement of
photosynthesis and transpiration additional 300% of the
elevation in transpiration rate to be added to the
photosynthesis rate in order to preserve constant WUE.
70 + 15% of 70 + 3* 15% of 70 = number of
photosynthesis units needed for CONSTANT WUE = 42
times more photosynthesis needed to preserve constant
WUE at 15 % more transpiration.
The duration of transpiration is the whole day, while the
photosynthesis is abundant only at day time. Therefore
the total daily transpiration of the transgenic plants is
68. 68
elevated MUCH more than the total photosynthesis.
The reports regarding the elevated WATER use efficiency
of the transgenic plants are not logical.
Every percent of elevation in transpiration levels is actually
more than 300 times more water per unit of dry weight.
The amount of CO2 molecules per single water is 70,
meaning that elevation in transpiration is serious to WUE.
For a single percent of added transpiration 300% of
photosynthesis is needed to keep constant WUE,
otherwise
WUE declines.
The cells which are full in water are larger in size, and
thus the time CO2 diffuses is longer. The longer time of
diffusion causes usually decline in photosynthesis rates in
highly irrigated plants.
The extent of photosynthesis addition of the transgenic
plants is incomparable to the extent of used water.
The improvement of 50% in photosynthesis is only in peak
irradiation hours and salt stressed plants.
To elevate WUE photosynthesis has to elevate 45 and
not 2 times.
NO ELEVATION IN WATER USE EFFICIENCY IS
EVIDENT EVEN IN THE TOP LIGHT and SALT STRESS
CONDITIONS,
when the aquaporin is the most affecting the plant.
69. 69
During normal conditions the differences in photosynthesis
are not large, however the difference in transpiration in
steady on ~16% from wt. Note that the plant transpires all
the day and not only at peak light hours.
Inconsistency of the results measured by gas-exchange
and whole plant scale measurement.
Figure 3 presents larger transpiration rates to transgenic
plants both at salt and normal irrigation. Figure 2 presents
transpiration results of whole plant. The whole plant
transpiration of transgenic plants is lower than WT.
Aquaporins improve water transport, and thus more
transpiration is expected.
HOW THE AUTHOR EXPLAINS THE INCONSISTENCY?
WHY IS PHOTOSYTHESIS ON NORMAL IRRIGATION
IS NOT PRESENTED?
The effect of the aquaporin is significant only when there
is water deficit. At normal irrigation, the photosynthesis
levels of Transgenic and wt plants should not differ
significantly. The reason of the aquaporin effect on
photosynthesis is that the photosynthesis is reduced by
high salt concentration during salt stress. Moreover, the
transgenic plants transpire more water and thus get faster
to drought stress. The addition of the aquaporin enhances
water transport, which is positively affecting
photosynthesis only at salt stress conditions, when the salt
levels start to block enzymatic processes.
CONSTANT LIGHT DURING THE GAS-EXCHANGE
EXPERIMENT
The light during the experiment was artificial and constant.
The reason for high transpiration at noon hours is
70. 71
temperature not irradiation.
Due to the artificial light, the photosynthesis was elevated
during the whole measurement period (morning=>noon),
which would have not been expected with natural light,
which is at PEAK only few hours a day. When light is less
intense, differences in photosynthesis rates are smaller.
The LIGHT levels was 1200uE which is stronger intensity
relative to real conditions. The light was TOO high in order
to intensify the photosynthesis rates of plants and
enhance the effect of the aquaporin on salt stress. LESS
LIGHT would have declined the TOTAL photosynthesis
and thus WUE would have been declined.
TOO HIGH :LIGHT is UNHONEST EXPERIMENT.
The real light is lower, not only to the top leaves but to the
rest of the plant. Most of the plant is at much lower light
levels (~200uE).
The differences in photosynthesis rates due to better
water transport during salt stress would have declined to
~5-10 % at maximum
relative to the reported 50%.
Note that the transpiration of the transgenic plants is
elevated at all conditions to more than 15%. At real
conditions,
the light levels are much lower, regarding the fact that at
morning and evening hours the light is very low intensity.
However, transpiration levels are related to temperature
and are not maximum, since the temperature of the air is
related to the real irradiation.
71. 71
CONSTANT LIGHT DURING EXPERIMENT IS A
FRAUD. THE TEMPERATURE OF THE AIR ALTERS
TRANSPIRATION,
BUT PHOTOSYNTHESIS IS KEPT MUCH MORE
CONSTANT DUE TO CONSTANT LIGHT.
THE PHOTOSYNTHESIS CURVE WAS NOT
PRESENTED MOST POSSIBLY DUE TO CONSTANT
LIGHT WHICH DOES NOT CORRELATE
TO THE CHANGING TRANSPIRATION.
NATURAL LIGHT DID NOT AFFECT
PHOTOSYNTHESIS, ONLY TRANSPIRATION.
Fig 5: Typical irradiation levels in a greenhouse with
natural light. UPPER LEAVES.
Rest of the plant, LESS than the following.
Average irradiation of morning: 450-500 uE
72. 72
Average irradiation of Noon : 750uE
The reported experiment : constant 1200uE from morning
to evening.
The publisher enhances photosynthesis, which is
enhanced by the aquaporin at SALT STRESS.
At real light conditions there is less photosynthesis due to
lower light levels.
The transpiration levels, however are kept according to
the air temperature and are influenced
by the changing irradiation levels.
The real light is HOT, and the artificial light is cold.
1200uE of SUNLIGHT would have elevated transpiration
significantly. 1200uE of artificial light, is constant,
from morning to noon and transpiration is related to air
temperature affecting on the whole plant. The effect of
irradiation
on transpiration is reduced due to the cold artificial light of
the measurement chamber.
The researcher accents photosynthesis and tries to hide
transpiration. In order to report POSITIVE WUE of
transgenic plants.
HOWEVER TO IMPROVE WUE, photosynthesis has to
rise by 45 times of the elevation of transpiration, during
the whole day
73. 71
Period due to mass differences of water and Carbon and
molar differences of transpiration and photosynthesis.
The SALT STRESS EXPERIMENT
Salt ACCUMULATION in plant force plants to grow BAD
on water with high salinity. The more plants transpire,
faster the accumulation of salt, and therefore the plants
live shorter lives until the plant death. Growth and fruit
count declines.
The transgenic plants are more sensitive to salt on the
longer run due to salt accumulation.
The addition of salt during the experiment is not a good
model for NATURAL field conditions. The fields which
absorb too much salt for plant growth should be replaced
with new soil. Drought stress is the real experiment to test
new genetically engineered plants. The drought stress is
predicted to be a failure for the transgenic plants carrying
the aquaporin which causes more transpiration and thus
the plants get stressed before the wt plants. Elevated
transpiration of the transgenic plants DECLINE WUE
causing drought sensitive plants or LARGER WATER
DEMAND.
The improvement of photosynthesis is ONLY at SALT
STRESS CONDITIONS WHICH ARE NOT REAL.
BETTER SOLUTION THAN GENETIC ENGINEERING IS
REPLACING THE SOIL.
Conclusions
1. WUE of the transgenic plants to decline since the
differences in photosynthesis rates are not high enough
relative to differences in transpiration.
75. 75
Leonid Asipov
11/15/2012 12:03:31 PM
Figure 1.
Stomatal opening in FC or KCI.
How possible that higher concentration of KCL open the
stomata and not closing? High concentration, water exits
the stomata and the stomata closes.
2. Why to couple 2 effects: solute concentration and light?
Light causes photosynthesis and thus decline in CO2
concentration.
When CO2 concentration is below threshold of stomata
opening, the stomata start to open.
Solute concentration is effect unrelated to light.
Cactuses create sugar during the day
Leonid Ldata
9/8/2011 3:39:39 PM
Cactuses create sugars from CO2 and water. During the
night, all the gas exchange takes place. The stomata
open, the O2 from the day irradiation diffuses to the air
and CO2 diffuses to the plant. The plant structure is low
76. 76
surface area/volume ratio and accumulates large amount
of CO2 and O2. At the morning stomata close and
irradiation start creating O, H and C from H2O and CO2.
The O2 and CO2 are accumulated in the plant leaves.
The sugar is enzymatically formed during the day, while
the basic components are created. As soon as stomata
are opened during the night, gas exchange takes place.
CO2 diffuses to the plant, where CO2 concentration is
low. O2 diffuses to the air. Without irradiation, the basic
components are becoming back to CO2 and H2O. During
the night only gas exchange takes place.
81. 81
באנרגייה ממחסור נושרים וכך , עצמו העלה לחיי אנרגייה
.
ל
-
דאטה
1122
שמורות הזכויות כל
There is no main growth control in plant :
transpiration is related to nutrient transport
Leonid Asipov
8/28/2011 9:41:40 AM
83. 81
Stomata
Stomata : CO2 Triggers
wep Leonid Ldata
1/23/2012 12:50:11 PM
Introduction
In this article we’ll show the relation between CO2
concentration in the air and the physiological parameters,
photosynthesis and transpiration. We’ve measured the
84. 84
stomata aperture at different CO2 concentrations and
came to interesting and conclusions regarding the
possible trigger for induction of changes in the stomata
aperture.
Responses of photosynthesis and transpiration to
changes in CO2 concentration
Elevation in CO2 concentration in the plant surrounding air
causes decrease in transpiration rate and increase in the
rate of carbon fixation. Decrease in CO2 concentration,
causes an exactly opposite effect.
Figure 1: Effect of changes in CO2 concentration on
photosynthesis and transpiration of Arabidopsis
85. 85
CO2 concentration has an effect on the transpiration rate in
light and in darkness (Figure 2), which suggests that
stomata reaction (which causes the changes in leaf
conductance to water) to CO2 is light independent.
Although in the light, the reaction is stronger.
Figure 2: Effect of changes in CO2 concentration on
the transpiration rate of Solanum lycopersicum in
light and darkness.
We’ve photographed stomata of Arabidopsis at different
CO2 concentrations of the air (figure 3). At 650 uL L-1 CO2
the stomata appear to be more closed than at 80 uL L-1.
This data suggest that CO2 may be the trigger for changes
in the stomata aperture.
86. 86
Figure 3: Stomata of Arabidopsis at 650 (left) and 80
(right) uL L-1 CO2.
What is the effect of light on transpiration and why in
the light, the response of the stomata is stronger than
in darkness (figure 2)?
It is stronger in light conditions because there is more
transpiration in light conditions, thus the effect of stomata
is elevated per unit of transpiration.
87. 87
Moreover that during the light period, the concentration of
CO2 in the leaf surroundings and the intercellular space is
lower than darkness due to photosynthesis. If CO2
concentration is the trigger, we would expect stronger
responses of stomata in light than in darkness. The
stomata would be more open and thus the transpiration
rate.
Can the concentration in the intercellular space be the
trigger for stomata induction?
In the following experiment (figure 4), transpiration,
photosynthesis and intercellular CO2 concentration was
measured during plants awakening from a period of
darkness.
We can see that up-regulation in the transpiration rate
happens not immediately with the light turning on (like the
photosynthesis), but after a period of time close to 20
minutes. During this period, the intercellular concentration
gets below a certain threshold. This experiment was
performed in low light levels of 20 µE.
88. 88
Figure 4: Photosynthesis, transpiration and
intercellular CO2 concentration during plants
awakening in low light (28uE).
Similar experiment was conducted in higher light levels of
300 µE. As we would have expected in the case that a
decline in the intercellular CO2 causes the stomata
response, at higher photosynthesis levels the delay
between the light turning on and transpiration response
should be shorter. This is exactly what we see in the
results (figure 5), the delay was shortened from 19 to 7
minutes.
89. 89
Figure 5: Photosynthesis, transpiration and
intercellular CO2 concentration during plants
awakening in high light (300 µE).
According to these experiments, intercellular CO2
concentration can be the trigger for the stomata aperture
induction. Further research, however, showed that it is
most likely not the intercellular but the concentration of
CO2 in the close surroundings of the guard cells, and it is
needed to relate to the gradient of CO2 from the stomatal
pore to the mesophyll cells which perform the carbon
fixation.
The problem with Ci (intercellular CO2) concentration
parameter is that it does not consider the gradient of CO2
between the stomatal pore and the photosynthesizing
mesophyll cells. The traditional Ci parameter conveys to
all the intercellular space the same concentration, which is
90. 91
not accurate, since the concentration of CO2 obviously has
a gradient between the stomatal pore and the
photosynthesizing mesophyl cells (figure 6).
Figure 6: The CO2 gradient in the intercellular space
In the following experiment we’ve measured
photosynthesis, transpiration and intercellular [CO2] during
stepwise elevation and decrease in the [CO2] in the leaf
surroundings (figure 7).
91. 91
Figure 7: The effect of stepwise CO2 concentration
changes on photosynthesis, transpiration and
intercellular CO2 concentration.
We see that after the elevation in CO2 concentration, there
is a decrease in the transpiration rate and intercellular
[CO2]. High CO2 levels cause stomata closure, and due to
high photosynthesis rate, the intercellular space becomes
less loaded with CO2. Despite the decrease in Ci to very
low levels, the stomata continue to be closed, against our
previous expectations (figures 4 and 5). Please notice that
the previous two experiments were conducted at 400 uL
L-1 CO2 and in the one shown in figure 7, the high CO2
point of this experiment was 600 uL L-1. At lower CO2
levels of 400 uL L-1, the gradient between the [CO2] of the
surroundings is less sharp than at 600 uL L-1, and thus at
400 uL L-1, Ci may represent more closely the [CO2] near
the guard cells. When the gradient is more sharp, the Ci
parameter represents less the [CO2] concentration near
the guard cells, which is closer to the [CO2] outside. The
93. 91
CO2 and Stomata
Leonid Asipov
11/15/2012 12:31:17 PM
The reason for the effect of CO2 on stomata aperture is
blocking of H2O near the plant cytoplasm. If CO2 is with
more concentration from outside, more chances are that
water molecules are diffusing from the cytoplasm to the
outer cell. If the concentration Is higher in the cytoplasm,
CO2 molecules are blocking near the cytoplasm
membrane and more molecules are diffusing from outside
to the cytoplasm. More CO2=> The stomata aperture
declines Less CO2=> Stomata aperture rises. The CO2 is
affecting the stomata without or with light.
Stomata do not affect photosynthesis, only
transpiration
94. 94
Leonid Ldata
9/11/2011 6:44:12 PM
The stomata does not affect photosynthesis,
only transpiration. Stomata aperture at any
state does not block photosynthesis,
only transpiration.
CO2 diffusion takes place in similar rates
despite stomata aperture. The proof is
photosynthesis rates at different water
irrigation levels. Watering plants does not
elevate photosynthesis, but elevates transpiration.
Watering usually causes lower photosynthesis
because longer time is needed for CO2
to diffuse to the leaf.
The next argument is the uncoupling of
photosynthesis and transpiration to CO2
changes. Photosynthesis changes almost
instantly (10-30 seconds) and transpiration
response is slow. Presuming, either the stomata
open or close fast as photosynthesis response,
or photosynthesis is not affected by stomata aperture.
Since transpiration is very affected by stomata,
and photosynthesis is not, most possibly
CO2 diffusion is fast and is not affected by
mechanical differences in stomata aperture.
95. 95
CO2 is trigger for stomata opening without any
biology
wep Leonid Ldata
2/21/2012 10:00:06 AM
96. 96
Stomatal response to CO2
Leonid Asipov
8/28/2011 10:34:57 AM
Stomata respond to CO2
As soon as the CO2 reach the stomata / other cell
surroundings the osmolarity rises and water exits the cell.
The cells are mechanical structures which open/close the
stomatal pore according to the pressure in the cell. High
pressure / the stomata is open,
low pressure- the stomata is closed.
When the concentration of CO2 lowers, water enters the
cell, and the stomata opens. The water pores are static
and have no function else than to be pores. The cells lose
water after CO2 elevation. To block the phenomenon
97. 97
there is structure which called stomata, which after water
transport to outside make a pore to close. Lower
concentrations of CO2 make the water enter the cell and
thus open the stomata.
CO2 is reactive molecule and affect the solute
concentration.
The stomata is acting as water sensor. Low water- the
stomata is closed.
CO2 makes water to exit the cells.
Water
The Role of the Difference Between Plasma and
Vacuole Membranes Water Conducting Properties : A
Model Approach
98. 98
Leonid Asipov
8/31/2009 7:50:21 PM
The model imitates cell submerged in a solution. As the
outer osmolarity changes, water flux is created, until
osmolarity in all the compartments reach equilibrium.
During this process, the submerged cell changes it’s
volume. The ratio in the volume between the cytoplasm
and the vacuole may change as well.
We’ll introduce a constant representing membrane’s
maximum water conductance properties per unit of time
(PP—Physical Permeability).
The water conducting properties of the both membranes
(PP), affect the maximum rate of water flux between the
cell compartments and the surroundings.
As experiments show, the difference in the PP of the
Vacuole membrane can get 100 or more times larger than
the Cytoplasm membrane.
99. 99
What is the functional logic of such difference ?
To answer that, we’ll simulate cell swelling and monitor the
osmolarity inside the cell compartments, at different
possible values of PP.
Vacuole PP = 1 Cytoplasm PP = 1(Click to see the movie
clip)
Vacuole PP = 10 Cytoplasm PP = 1 (Click to see the
movie clip)
100. 111
The results of both simulations are presented in the
following chart.
