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Development of Disease Resistant, Lea (low Erucic Acid) Quality BARI Sarisha-14 Through Marker-assisted Backcrossing (2014-2015)

M. M. Karim
Biotechnology Division, BARI, Joydebpur, Gazipur

M. A. Y. Akhond
Biotechnology Division, BARI, Joydebpur, Gazipur

M. A. L. Akanda
Biotechnology Division, BARI, Joydebpur, Gazipur

M.S. Hossain
Biotechnology Division, BARI, Joydebpur, Gazipur

M. R. Kabir
Biotechnology Division, BARI, Joydebpur, Gazipur

M. S. Hossain
Biotechnology Division, BARI, Joydebpur, Gazipur

Abstract/ Executive Summary:

Short-duration Brassica rapa cultivating in the country has no low erucic acid (LEA) cultivars. In order to develop low erucic acid mustard cultivars, high erucic acid (HEA) mustard cultivars were interspecifically crossed as recurrent parents to a canola quality rapeseed. In the advanced backcross, homozygous e1e1 type lines were developed through marker-assisted selection. Fatty acid composition in seeds revealed very low erucic (0.3 - 0.8%), high oleic (53.0 - 67.9%), and moderate linoleic (9.7 - 17.8%) acid contents, which belong to the zero erucic acid class. The developed line also contained Fusarium wilt-resistant loci as well as short-duration quality. Others yield contributing characters also found satisfactory in some advanced lines compare to cultivated short duration B. rapa, BARI sarisha-14. So, promising lines will contribute to Fusarium resistant, Low erucic acid (LEA) quality short duration rapeseed breeding.

Keywords: Homogyzous, Marker assisted backcrossing, Mustard

Location:

Start Date: 00-00-2014

End Date: 00-00-2015

Program Area: Variety and Species

Commodity: Mustard

Objectives:

Fusarium oxysporum resistant gene of B. rapa, FocBr1 has been identified by Shimizu et al. 2014. Furthermore, genetic mapping of a Fusarium wilt resistance gene in Brassica oleracea also reported (Pu et al. 2012). So, Fusarium wilt resistance and LEA quality gene will be introgressed through MAS into BARI sarisha-14.

Methodology:

Backcross progenies of BARI sarisha-14 with canola cultivar were evaluated for disease resistance, early flowering and LEA quality after marker assisted backcrossing. Fourteen yellow seeded BC3S2?hetero plants, containing functional and non-functional both BnFAE1.1 allele (E1e1) was grown in summer 2014 at Biotechnology division greenhouse. Marker-assisted selection (MAS) was done to screen yellow seeded non-functional allele type (e1e1) BC3S2 generation.

Fatty acid composition analysis was performed using the BC₂S₃ - BC₃S₄ seeds from the plants predicted to be homozygous for BnFAE1.1 along with the parental lines. Identification and quantification of fatty acid compounds were accomplished using capillary gas chromatography with flame ionization detection (GC–FID).

In winter 2014-15, yellow seeded mutant BC3S3 generation and other brown seeded mutant BC2S3 and BC3S2 were evaluated for earliness and yield in Oilseed Research Centre (ORC), Joydebpur, Gazipur experimental field. Short duration plants from advanced lines were selected through selfing. Seed was sown on 13^th November 2014. Row to row and plant to plant distance were maintained 40cm×10cm after thinning. Plot to Plot distances were 70 cm and row to row distances were maintained one meter. In case of field experiments, there were six advances low erucic acid (LEA) lines as a treatment along with Tori-7 and BARI sarisha-14 (HEA) as a check variety.

Source of Information: Annual Research Report 2014-2015, Biotechnology Division, BARI, Joydebpur, Gazipur

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

By using the molecular markers for Fusarium wilt resistance, It was found that the recurrent parents (Tori-7 and BARI sarisha 14) and donor parent (Kirariboshi natane) contained A-genome specific Fusarium wilt resistant loci which were used to develop Low Erucic Acid (LEA) quality short duration B. rapa. As both parents contained Fusarium wilt resistant allele so, it might transfer their advanced progeny also. In BC3S2 generation, some plants contained both functional and non-functional alleles. Through selfing of those hetero-type plants, three types of allele combinations were found. After digestion with restriction enzymes, three types of band patterns which correspond the E1E1, e1e1, and E1e1 were found. Accordingly, in the backcrossed progeny derived from Kirariboshi × Tori-7, the PCR product of recurrent B. rapa plants (E1E1) contained a recognition site to produce ca. 850bp plus 22bp bands after digestion (Fig.2), whereas the PCR product of heterozygous plants (E1e1) produced double bands, one of which was a non-cut band (872bp) corresponding to mutant allele (e1) and the other was the 850bp band (E1). Finally, heterozygous BC3S2 plants (E1e1) were self-pollinated so that the non-functional e1 allele was fixed in the B. rapa genome, where e1 allele was incorporated via homologous recombination of A-genome of B. rapa and B. napus during the backcrossing process. All three genotypes (E1E1, E1e1 and e1e1) produced yellow seeds but different in % erucic acid (Fig.3). The parental B. napus cv. Kirariboshi has 1.2% erucic, 63.1% oleic, and 15.1% linoleic acids in seeds (Table 1). The B. rapa parental genotypes, Tori-7 and BARI sarisha-14, contain 45.4% and 47.1% erucic, 15.6% and 14.9% oleic, and 12.2% and 13.9% linoleic acids in seeds, respectively. In the backcrossed progenies, the erucic acid content decreased in a dose-dependent manner on the mutant type allele; 33.9 - 43.6 % erucic acid in wild type plants (E1E1), 19.1 -19.8 % erucic in heterozygotes (E1e1), and 0.3 - 0.8 % erucic acids in mutants (e1e1), respectively. In the homozygous e1e1 type lines, fatty acid composition in seeds revealed very low erucic (0.3 - 0.8%), high oleic (53.0 - 67.9%), and moderate linoleic (9.7 - 17.8%) acid contents, which belong to the zero erucic acid class. Yield, yield contributing characters and maturity data of different low erucic acid (LEA) containing advanced backcross progenies along with their recurrent parents were presented. BARI sarisha-14 produced the highest 1000 grain weight (3.8g) followed by T7; BC3S3 (11-1-3-7-14) B14 (3.4g). Tobin is a canola quality Canadian variety that produced the highest yield/5plant (9.1 g) followed by T4; BC2S3 (3-6-4-7) B14, T3; BC2S3 (3-6-4-3) B14. BARI sarisha-14 showed the earliest maturity followed by Tori-7 and T7; BC3S3 (11-1-3-7-14) B14. Plant height, number of branches per plant, flowering time, Alternaria blight disease and % oil content data of different low erucic acid (LEA) containing advanced backcross progenies along with their recurrent parents were recorded. Fatty acid composition in recurrent B. rapa parents (Tori-7, BARI sarisha-14), donor B. napus parent (Kirariboshi) and advanced backcross lines

Knowledge Management: Report/Proceedings