chlorophyll mutation

1. 1
Suresh Yadav
I.D No. - PALM 6005

2. FLOW OF PRESENTATION:-
INTRODUCTION :-
COMPARISION BETWEEN M1 AND M2 WITH F1 , F2 POPULATION :-
Identification of Mutant Plants in M1 Generation :-
PROCEDURES FOR SELECTING MUTANTS :-
CASE STUDIES :-
CONCLUSION :-
SUMMARY :- 2

3. INTRODUCTION :-
 Role of Mutagenic Treatments:-
 Genetic features of M1 and M2 Population:- Differ from F1 And F2 ?
Most of The Induce Mutations are recessive.
 M1 Population :-
 Mutation Rate Per Cell :-
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5.  Identification of Mutant Plants in M1 Generation:-
 Difficult to Identify ?
M2 Population :- Proporation of Mutant (a’a’) to wild type(AA) less than that of
Segregating in Mendalian Fashion.
Procedures for Selecting Plants :-
Growing out the M1 Population:-
Guide line to be follow :-
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7. Procedures for Selecting Mutants :-
 M1 Plants have higher out crossing tendancy than non treated plants
?
 Effect ?
Harvesting and growing of M2 seed:-
 Basically two types:-
1. M1 spike progeny Method :-
 Developed by Stadler (1928)
Usually 10-20 plants grown out from seed of Individual M1 spikes.
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8.  1. M1-spike progeny Method :-
• Segregation Frequency :- The Frequency of mutants per progeny.
• Segregation ratio :- The ratio of a trait Segregating according to its genetic
control. (e.g. single gene controlled trait ratio 3:1).
• Factor affecting Segregation Frequency :-
2. One- plant- one –grain Method:-
 Also known as “ one spike-one-grain method.”
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9. EFFECT OF CHEMICAL AND PHYSICAL MUTAGEN IN M1 GENERATION AND
CHLOROPHYLL MUTATIONS IN SOYBEAN
(GLYCINE MAX L. MERRILL.)
MATERIALS AND METHODS:-
Dry seeds of “MACS 450” exposed to gamma rays.
 Followed Subjected to EMS (0.2% and 0.4%)
• Combination of both (50Gy + 0.2% EMS, 50Gy + 0.4% EMS, 100Gy + 0.2%EMS and
100Gy + 0.4% EMS).
In another experiment seeds irradiated with gamma rays at 50Gy were treated
with SA(0.003 %and 0.005 % w/v NaN3).
 for combination treatments of gamma rays and SA (50Gy + 0.003 % NaN3,50Gy +
0.005 % NaN3).
( Dig ambar Ahire*1 and Sanjay Auti,2015.)

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10. MATERIALS AND METHODS:-
• Untreated soybean seeds presoaked in distilled water for duration
viz,
• 12 hrs, 16 hrs and 20 hrs.
• The presoaked seeds treated with 0.8% (v/v) EMS solution.
• The 0.8% EMS concentration was selected on the basis of (LD 50)
• lethal dose from pre experiments.
• washed in tap water for 2 hrs.
Treated seeds sown along with control under RCBD with 3 replications.
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11. Observations were recorded:-
• Survival (%).
• plant height
• pods/ plant
• seeds/ plant
• Seeds/pod
• Chlorophyll mutations were scored emergence of cotyledons in M2 population.
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15. RESULTS AND DISCUSSION :-
• higher dose of 100Gy +0.4% EMS effective that induced 36.4 %
reduction in plant ht. & 29.2% seed sterility.
• The highest conc. of chemical mutagens inhibited morphological
development of seeds and seed viability.
• Combination treatments 50 Gy + 0.005% SA caused more damage.
• EMS treatment injury less in 12 hrs presoaked in water.
• Reduction in plant height doe to drop in auxin level, chromosomal
aberrations, decline of assimilation mechanism.
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16. CONTINUE…
• Treatments,of gamma rays and NaN3 (50 Gy + 0.005% SA) exhibited
maximum seed sterility (33.3%).
• 100Gy +0.4% EMS was exhibited maximum reduction in height (36.4%)
and
• pod /plant (35.3) (Table1).
• Studies in M2 generation:-
• Frequency and spectrum of chlorophyll mutations:
• chlorophyll mutants albina, xantha, chlorina, viridis and sectorials
(chimera).
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17. CONTINUE:-
• Among gamma rays and EMS combination treatments 50Gy +0.4%
EMS and 100Gy+0.4% EMS induced broader spectrum.
• (0.49-2.91%) In M2 plant (Table 2).
• Among all the mutagenic treatments the range of chlorophyll was
from 0.15 - 4.50%.
• Treatment EMS 0.8% to 16hrs presoaked seeds was the most effective
one (Table 4.2).
• Viridis appeared more than others ?.
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18. Origin of cholorophyll deficiency :-
• Due to mutations in genes.
• Responsible for synthesis of photosynthetic pigments.
• Chlorophyll deficient mutants lack well-defined grana structure in
chloroplasts.
• :- CONCLUSION :-
• Combination of mutagens are more effective than individual
treatment.
• Higher dose of 100Gy +0.4% EMS was more effective.
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19. :- CONCLUSION :-
• Frequency of chlorophyll mutations increased with higher
concentration of EMS (0.4%) with 50 and 100 Gy gamma rays dose in
combination treatment.
• Viridis appear in maximum % and albina in less percentage in all
treatments over other types of chlorophyll mutations.
• This help in find out type and effective concentration of mutagen.
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20. CASE STUDIES :- 2
• Effects of mutagen on chlorophyll mutation in horse gram
[macrotyloma uniflorum (Lam) Verdcourt]
• MATERIALS AND METHODS :-
• (9-12 % moisture) and healthy seeds of Horse gram [Macrotyloma
uniflorum (Lam) Verdcourt].
• Three chemical mutagen
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21. CONTINUE:-
Mutagen CONC. (%) CONC. (%) CONC. (%)
EMS 0.1 0.2 0.3
SA 0.05 0.1 0.15
NMU 0.4 0.6 0.8
EMS (Ethyl Methane Sulfonate), SA (Sodium
Azide) and NMU (N-Nitroso-N-Methyl Urea)
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22. CONTINUE:-
• Treated seed presoaked in distilled water for 4 hrs at room temp.
followed by six hours in mutagenic solutions.
• Thoroughly wash in running water for 4 hrs.
• 300 seeds from each treatment sown in RBD with 3 replications with
control for rising M1.
• Seeds of 25 randomly selected. used for rising M2 generation and
chlorophyll mutations recorded when seedlings were 10-15 days old.
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23. RESULTS AND DISCUSSION :-
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24. RESULTS AND DISCUSSION :-
• 50% reduction in seed germination at 0.4% of EMS, 0.2% of SA and 1.0% of
NMU.
• Based on Lethality 0.3% of EMS (2.23%), 0.15% of SA (1.59%) and at 0.8%
of NMU (1.28%).
• In this experiment, no distinct mutant’s plants were found regarding yield
components which died permanently.
•
• Mutation frequency showed gradually increased with the increasing
concentration of mutagens.
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25. RESULTS AND DISCUSSION :-
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26. RESULTS AND DISCUSSION :-
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27. RESULTS AND DISCUSSION :-
• Among mutagens, 0.3 % EMS proved efficient in inducing mutations
with highest frequency (Table1).
• Mutagenic effectiveness as determined on M2 plant basis was
NMU>SA>EMS.
• Physiological mutants early and late maturity observed in mutagenic
treatments.
• The maximum number of early and late maturity mutant was
observed at 0.2 and 0.3% of EMS.
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28. SUMMARY :-
• Genetic features of M1 and M2 Population.
• Most of The Induce Mutations are recessive.
• Identification of Mutant Plants in M1 Generation.
• Procedures for Selecting Plants :-
• EFFECT OF CHEMICAL AND PHYSICAL MUTAGEN IN M1 GENERATION AND CHLOROPHYLL MUTATIONS
IN SOYBEAN (GLYCINE MAX L. MERRILL.)
• Effects of mutagen on chlorophyll mutation in horse gram [macrotyloma uniflorum (Lam) Verdcourt]
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