Exploitation of in vitro culture techniques for the development of new genetic combinations in sugarcane

Transcription

1 Exploitation of in vitro culture techniques for the development of new genetic combinations in sugarcane Dr. Imtiaz A. Khan Pr. Scientist / PI sugarcane and molecular marker group NIA-2010 NIA-2012 NIA-2011

2 What is in vitro culture techniques? How about some definitions General Definition The application of technology to improve a biological organism Detailed Definition The application of the technology to modify the biological function of an organism by genes modification

3 What controls this natural variation? Allelic differences at genes control a specific trait Definitions are needed for this statement: Gene - a piece of DNA that controls the expression of a trait Allele - the alternate forms of a gene

4 Central Dogma of Molecular Genetics (The guiding principle that controls trait expression) Protein Trait (or phenotype) Translation Seed shape DNA (gene) Transcription RNA Plant height

5 Rational of applying biotechnological techniques for sugarcane improvement 4. Chromosomal instability 3. Scares arrangement of artificial flowering 2. Flower with 98% sterility 1. Non or sporadic flowering

6 Sugarcane improvement through biotechnological techniques

7 Callus culture studies C) Regeneration A) Explant D) Rooted plantlets B) Callus E) Plantlets in earthen pot F)Plants in the field

8 MICROPROPAGATION IN SUGARCANE

9 Explant of sugarcane Callus induction type A In vitro mutagenesis studies Nonregenerablecallus TypeB

10 Parent Somaclone

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12 Plantlets regenerated through callus culture

13 Plantlets regenerated through in vitro mutagenesis

14 Somaclones Parent (NIA-2004)

15 Plantlets Regenerated Through Direct Regeneration Method

16 Variability observed in the mutants of CPF Characters Parent Mutants (range) % of variation Stalk/stool (no.) to Cane length (cm) to Cane thickness (cm) to 1.55 Cane weight (kg) to Cane yield (t/ ha) to 7.36 CCS % to Fiber % to 1.81 Sugar yield (t/ha) to 32.09

17 Variability observed in the somaclone of NIA Characters Parent Mutants (range) % of variation Stalk/stool (no.) to Cane length (cm) to Cane thickness (cm) to Cane weight (kg) to Cane yield (t/ ha) to CCS % to Fiber % to Sugar yield (t/ha) to 61.77

18 Variability observed in the somaclones of CP Characters Parent Mutants (range) % of variation Stalk/stool (no.) to Cane length (cm) to 9.65 Cane thickness (cm) to 4.06 Cane weight (kg) to 9.18 Cane yield (t/ ha) to CCS % to Fiber % to Sugar yield (t/ha) to 54.03

19 DIRECT REGENERATION IN SUGARCANE

20 DIRECT REGENERATION IN SUGARCANE

21 Variability observed in the direct regenerants of Thatta-10 Characters Parent Variants (Th-10) (range) % of variation Stalk/stool (no.) to Cane length (cm) to 9.52 Cane thickness (cm) to 4.00 Cane weight (kg) to 8.88 Cane yield (t/ ha) to CCS % to 6.17 Fiber % to Sugar yield (t/ha) to 27.72

22 Variability observed in the direct regenerants of SPSG-26 Characters Parent (SPSG26) Variants (range) % of variation Stalk/stool (no.) to 125 Cane length (cm) to Cane thickness (cm) to Cane weight (kg) to Cane yield (t/ ha) to CCS % to Fiber % to Sugar yield (t/ha) to 58.80

23 Variability observed in the direct regenerants of L-116 Characters Parent (L116) Variants (range) % of variation Stalk/stool (no.) to Cane length (cm) to 7.93 Cane thickness (cm) to 0.93 Cane weight (kg) to 7.14 Cane yield (t/ ha) to Sucrose % to Fiber % to 6.12 Sugar yield (t/ha) to 53.36

24 SUGAR MILLS CO-ORDINATED TRIAL Mehran sugar Mills Farm, Tando Allahyar Matiari Sugar Mills Farm, Matiari Faran Sugar Mills Farm, Sheikh Berkio Pangrio Sugar Mills Farm, Pangrio Mirpurkhas Sugar Mills Habib Sugar Mills, Nawabshah Ranipur Sugar Mills Genotype Planted NIA-2004 NIA-2010 NIA /P5 CPF-237/SPF-234 Thatta-10

25 Two years Pooled data of Sugar Mills Sugar Mills Coordinated trial Clones / Locations Stalk Cane / length stool (cm) (Nos) Cane girth (cm) Weight/ stool (kg) Cane yield (t/ha) CCS % NIA820819/P5 5.8ab a 2.57 c 5.67 b a a NIA bc b 2.38 d 5.23 b 137.0a b NIA c c 2.43 d 4.55 c ab b CPF a 226.6b c 3.03 a 5.31 b b Thatta c c 2.75 b 6.68 a ab b c

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29 NIA-2011

30 NIA-2010

31 Visit of sugarcane Expert from Sri lanka and AARI Faisalabad

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34 Figure 1. TRAP profile of sugarcane genotype using surose synthase; M=DNA marker, 1= AEC , 2= GT-11, 3= AEC92-105, 4= AEC , 5= Thatta-10, 6= AEC82-223, 7= AEC , 8= NIA-2004,9= AEC86-328, 10= L116, B= Blank

35 Figure 2. STS profile of sugarcane genotype using DREB sequence; M=DNA marker, 1= AEC , 2= GT-11, 3= AEC92-105, 4= AEC , 5= Thatta-10, 6= AEC82-223, 7= AEC , 8= NIA-2004, 9= AEC86-328, 10= L116, B= Blank

36 Genetic variability in sugarcane biotechnological techniques Cane weight (kg) MB IVM DR CC MB Cane thickness (cm) Stalk/stool (no.) developed CC DR IVM through

37 Genetic variability in sugarcane biotechnological techniques Cane length (cm) developed through Cane yield (t/ ha) Cane yield (t/ ha) 0 MB CC DR Cane length (cm) IVM

38 Genetic variability in sugarcane biotechnological techniques developed through Sucrose % CCS % Fib % Sugar yield (t/ha) Sugar yield (t/ha) Fib % CCS % Sucrose % 10 0 MB CC DR IVM

39 Accession number Status NIA-2013 Sugar Mills coordinated Trial NIA-2012 Sugar Mills coordinated Trial NIA-2014 Sugar Mills coordinated trial NIA-03/2011 BL4-P70 (somaclone) 15 promising clones are in different stages of evaluation Will be advanced to zonal trial next year Sugar Mills coordinated trial In different stages of evaluation

40 Thanks for patience listening Your Questions Please