Induction of somaclonal variation in bajra (Pennisetum glaucum (L.) R. Br.) x napier grass (Pennisetum purpureum Schum.) hybrid through tissue culture
Loading...
Date
2021
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
Punjab Agricultural University, Ludhiana
Abstract
The present study was conducted with a major objective of standardization of protocol for callus
induction, shoot and root induction and elongation in Bajra × Napier grass hybrids and parents and
regeneration of plants from long maintained calli and further molecular, morphological and
biochemical characterization of regenerated plants. Optimal medium identified for callus
induction and its sub culture was MS + 2,4-D (2 mg/l) + BAP (0.5 mg/l). For shoot regeneration
and elongation MS + BAP (2 mg/l) + NAA (0.1 mg/l) was found to be most effective. Optimal
medium for root induction was ½ MS containing 25 ml stock I, 10 ml of stock II and stock III
each and remaining supplements according to 1 litre medium composition. Calli were sub cultured
at 21 days interval for 6 months. Maximum callus induction was 75.60% in PCB 164 whereas
Capricorn showed least callus response towards callusing (12.64). BNH 11 produced maximum
(50%) embryogenic callus whereas minimum embryogenic callus was observed in PCB 164
(11.29%). Plantlets were then regenerated from long maintained calli and hardening was
performed for 7 days and plants transferred to field in and augmented design. PCB 164 showed
maximum per cent of shoot and root induction with values 33.33 and 100% respectively. Albino
plants were only seen for PBN 346 (12.04%). DNA was isolated from field grown regenerated
plants and amplified using 17 SSR primers of Pearl millet. Somaclones were identified on the
basis of change in banding pattern from parental checks. Total of 32 somaclones were identified
based upon 5 primers viz. Xcump010, Xctm10, Psmp2085, Psmp2008 and Xpsmp2248 that were
polymorphic. PBN 346-7 showed change in banding pattern for maximum number (3) of markers.
ANOVA analysis revealed significant mean sum of squares for control versus augmented.
Correlation was calculated using Pearson coefficient method and significant correlation was found
among different traits. Principle component analysis (PCA) revealed that PC1 and PC 2 accounted
for 96.9% variability. Clustering of 103 genotypes was performed using Euclidean dissimilarity
index on ‘R’ software that revealed high variation among PBN 346 and BNH 11 genotypes. Based
on significant difference of value of different traits from their respective checks; Somaclones
identified on the basis of NOL, NOT, SG and IVDMD were 34, 51, 33 and 2 respectively in case
of PBN 346 and 2, 2, 3 and 6 in case of BNH 11 somaclones. Somaclones showing maximum
values for traits NOL, NOT, SG and IVDMD were PBN 346-56 (237.08), PBN 346-48 (54.83),
PBN 346-22 (10.27 mm) and BNH 11-98 (53.71%) respectively. Among somaclones that were
identified based on molecular data, number of somaclones showing significantly higher values
than respective checks for specific traits were 12, 17, 10, 3 for NOL, NOT, SG, and IVDMD
respectively. Common somaclones that were identified as somaclones on both molecular and
phenotypic basis were 23 in number.
Description
Keywords
Citation
Sidhu, Navjot Singh (2022). Induction of somaclonal variation in bajra (Pennisetum glaucum (L.) R. Br.) x napier grass (Pennisetum purpureum Schum.) hybrid through tissue culture (Unpublished M.Sc. thesis). Punjab Agricultural University, Ludhiana, Punjab, India.