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Selection of Microspore Derived Embryo Genotypes Based on Fatty Acid Pattern Analysis in Rapeseed

F Shafrin
MS Student
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh, Bangladesh

U K Nath
Professor
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh, Bangladesh

M S Alam
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh, Bangladesh

D Akhter
Department of Genetics and Plant Breeding, Sylhet Agricultural University, Sylhet, Bangladesh

Abstract/ Executive Summary:

In mustard (Brassica campestris L.), microspore culture is frequently applied for immediate regeneration of homozygous doubled haploid (DH) plants. In this study, our prudent concern is to develop a simple and efficient selection protocol to develop doubled haploid, based on fatty acid pattern present in MDEs. The feasibility of the protocol was carried out using Agrani, Safal, BARIsarisha-2, TORI-7 & BINAsarisha-6 as source of explants. Microspores of the 5 selected genotypes were cultured in liquid NLN medium and established embryos were sub-cultured in MS solid medium. Single cotyledon was dissected from corresponding MDEs of each genotype, under aseptic conditions. Fatty acid pattern analysis was conducted through Gas Liquid Chromatograghy (GLC). Thus, after analysis, it was found that all the genotypes had conspicuous effect on all examined fatty acids. Among them TORI-7 possessed 47.29% erucic acid (22:1) in MDE oil. Polyunsaturated fatty acid (PUFA) content ranged 20-32% for all the genotypes included in microspore culture. Safal exhibited 36% oleic acid (18:1), which could be recommended for cultivation as source of fried oil. BINA sarisha-6 given 9% saturated fatty acid (SFA). Correlation coeffients among the fatty acids revealed that only SFA had significant (-0.911) negative effect on erucic acid content. SFA and oleic acid were highly heritable traits (93-97%). PUFA had the highest direct effect on erucic acid content.

Keywords: Microspore, MDE, Fatty acids

Location: Laboratory of Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh, Bangladesh

Start Date: 00-10-2009

End Date: 00-02-2010

Program Area: Variety and Species

Commodity: Mustard

Objectives:

To develop a simple and efficient selection protocol to develop doubled haploid, based on fatty acid pattern present in MDEs.

Methodology:

Five rapeseed and mustard genotype viz. Agrani, Safal, BARIsarisha-2, TORI-7 and BINAsarisha-6 were used as source of explants for conducting the research for the production of doubled haploid through microspore culture following the protocol described.Cotyledon dissection: From fully developed embryos single cotyledon was dissected for fatty acid analysis . Selection in vitro for oil seed quality traits is possible in segregating populations of MDEs . Fatty acid composition was determined by gas liquid chromatography (GLC) from single cotyledon, dissected from MDEs. Fatty acid analysis by GLC (Gas liquid chromatography): Fatty acid composition was determined by gas liquid chromatography (GLC) from single cotyledons, dissected from MDEs. The rest of the MDEs were maintained in vitro and regenerated to plantlets. Unequivocal selections for different fatty acids were possible in segregating MDE population. However, the dissected cotyledon was grounded in a 1ml glass vials by adding 200 µl iso-octane- isopropanol solution (9:1). The mixture was air dried and transmethylated by adding 200 µl of 5% NaHSO₄ solution. Subsequently, 300µl isooctane was added to the sample shaking them on a Vortex. Pipetted out ca. 180 µl of the upper phase after 5 min centrifugation at 150 x g (1000 rpm) was transferred into septum vial, and following injection (ca. 2 µl) into the GLC (PERKIN ELMER 8420, San Jose, CA, USA) equipped with auto sampler. The gas chromatography was equipped with a fused silicon capillary column FFAP (Mecherey & Nagel GmbH+ Co. kg, Duren, Germany) 25 m x .25 mm i.d. (0.1 µm film thickness). The column temperature was 210^o C and carrier gas was 150 kPa H₂ at the split rate of 1:70, and injection/ detector temperature was 230^o C. Individual fatty acids, such as palmitic, stearic, oleic, linolenic, eicosenoic, erucic acid were determined by GLC and expressed as % of total fatty acids.

Source of Information: Int. J. BioRes. 2(12): 05-07, 2010

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

It was found that all the genotypes had conspicuous effect on all examined fatty acids. Among them TORI-7 possessed 47.29% erucic acid (22:1) in MDE oil. Polyunsaturated fatty acid (PUFA) content ranged 20-32% for all the genotypes included in microspore culture. Safal exhibited 36% oleic acid (18:1), which could be recommended for cultivation as source of fried oil. BINA sarisha-6 given 9% saturated fatty acid (SFA). Correlation coeffients among the fatty acids revealed that only SFA had significant (-0.911) negative effect on erucic acid content. SFA and oleic acid were highly heritable traits (93-97%). PUFA had the highest direct effect on erucic acid content.

Knowledge Management: Journal