Inheritance of siliqua locule number and seed coat colour in Brassica juncea

The inheritance of siliqua locule number and seed coat colour in Brassica juncea was investigated, using three lines each of tetralocular brown seeded and bilocular yellow seeded. Three crosses of tetralocular brown seeded × bilocular yellow seeded lines were attempted and their F₁, F₂ and backcross generations were examined for segregation of these two traits. Brown seed colour and bilocular siliqua characters were found to be dominant over yellow seed and tetralocular siliqua, respectively. Chi‐square tests indicated that each trait is controlled by different sets of duplicate pairs of genes. Bilocular siliquae or brown seeds can result from the presence of either of two dominant alleles, whereas tetralocular siliquae or yellow seeds are produced when alleles at both loci are recessive. A joint segregation analysis of F₂ data indicated that the genes governing siliqua locule number and seed colour were inherited independently.     

Development of two new cytoplasmic male-sterile lines in Brassica juncea through wide hybridization

Two new cytoplasmic male-sterile (CMS) lines have been developed in Brassica juncea using the bridgecross hybrids (Diplotaxis erucoides×Brassica campestris)×B. juncea and (Diplotaxis berthautii×B. campestris)×B. juncea. These were backcrossed ×ve times with pollen of B. juncea. The CMS line (D. erucoides) B. juncea segregated into tall and short true breeding types; both resembled the cultivar B. juncea in vegetative and floral morphology and in cytology, except for a greater number of secondary branches and smaller anthers with empty sterile pollen in the CMS line. Female fertility was as good in the CMS line as in the cultivar. The other CMS line (D. berthautii) B. juncea resembled the cultivar in vegetative morphology and cytology. Four true breeding floral types were isolated as follows: (1) smaller and indehiscent anthers containing empty, sterile pollen, (2) all six stamens petaloid, (3) one petaloid stamen and five stamens antherless, and (4) apetalous flowers with all six stamens antherless.     

Production of yellow-seeded Brassica napus through interspecific crosses

Yellow‐seeded Brassica napus was developed from interspecific crosses between yellow‐seeded Brassica rapa var.‘yellow sarson’ (AA), black‐seeded Brassica alboglabra (CC), yellow‐seeded Brassica carinata (Bbcc) and black‐seeded B. napus (AACC). Three different interspecific crossing approaches were undertaken. Approaches 1 and 2 were designed directly to develop yellow‐seeded B. napus while approach 3 was designed to produce a yellow‐seeded CC genome species. Approaches 1 and 2 differed in the steps taken after trigenomic interspecific hybrids (ABC) were generated from B. carinata×B. rapa crosses. The aim of approach 1 was to transfer the yellow seed colour genes from the A to the C genome as an intermediate step in developing yellow‐seeded B. napus. For this purpose, the ABC hybrids were crossed with black‐seeded B. napus and the three‐way interspecific hybrids were self‐pollinated for a number of generations. The F₇ generation resulted in the yellowish‐brown‐seeded B. napus line, No. 06. Crossing this line with the B. napus line No. 01, resynthesized from a black‐seeded B. alboglabra x B. rapa var.‘yellow sarson’ cross (containing the yellow seed colour genes in its AA genome), yielded yellow‐seeded B. napus. This result indicated that the yellow seed colour genes were transferred from the A to the C genome in the yellowish‐brown seed colour line No. 06. In approach 2, trigenomic diploids (AABBCC) were generated from the above‐mentioned trigenomic haploids (ABC). The seed colour of the trigenomic diploid was brown, in contrast to the yellow seed colour of the parental species. Trigenomic diploids were crossed with the resynthesized B. napus line No. 01 to eliminate the B genome chromosomes, and to develop yellow‐seeded B. napus with the AA genome of ‘yellow sarson’ and the CC genome of B. carinata with yellow seed colour genes. This interspecific cross failed to generate any yellow‐seeded B. napus. Approach 3 was to develop yellow‐seeded CC genome species from B. alboglabra×B. carinata crosses. It was possible to obtain a yellowish‐brown seeded B. alboglabra, but crossing this B. alboglabra with B. rapa var.‘yellow sarson’ failed to produce yellow seed in the resynthesized B. napus. The results of approaches 2 and 3 demonstrated that yellow‐seeded B. napus cannot be developed by combining the yellow seed colour genes of the CC genome of yellow‐seeded B. carinata and the AA genome of ‘yellow sarson’.     

Development of self-incompatible Brassica napus: (I) introgression of S-alleles from Brassica oleracea through interspecific hybridization

Cultivars in Brassica napus var. oleifera, a self‐pollinating, self‐compatible species, have traditionally been developed as open‐pollinated lines or populations. Significant yield gains in this species have been realized through the exploitation of heterosis. Commercial hybrid production has been possible as a result of the development of a number of pollination control systems. Self‐incompatibility was transferred from B. oleracea var. italica to B. napus var. oleifera through interspecific hybridization. The response to interspecific pollination, as measured by pod elongation and initial stages of ovule development, was genotype dependent, and two highly responsive B. napus genotypes were identified. Embryo rescue was used to produce the interspecific hybrids. Isoelectric focusing of stigma proteins was used to identify S‐alleles in the interspecific hybrids to facilitate backcrossing. Segregation of the S‐locus through a series of back‐crosses to B. napus was complicated by aneuploidy; however, the S‐locus was found to segregate as a single gene. Usefulness of B. oleracea as a source of S‐alleles for pollination control in B. napus is discussed.     

Plant Regeneration from the Hybridization of Brassica juncea and B. napus Through Embryo Culture

Crosses were made to produce interspecific hybrids between Brassica napus × B. juncea and their reciprocals with the aid of embryo culture techniques. A better response of hybrid embryo culture was obtained from two cross combinations of B. juncea × B. napus (Ames 24521 × Huyou 15 and Vittasso × Zheshuang 72) than from their reciprocals. Embryo culture was more effective in terms of plant regeneration when embryos were cultured in vitro at 15 days after pollination (DAP), while more calli were initiated when embryos were excised and cultured at 10 DAP. A better response was observed on the MS medium with 0.3 mg l^?1 naphthylacetic acid (NAA) + 1.5 mg l^?1 6‐benzylaminopurine (BAP) and with 0.3 mg l^?1 NAA + 2.0 mg l^?1 BAP. Callus formation and plant regeneration on these two media reached 55.43 and 26.65 %, and 66.98 and 24.61 %, respectively.     

Development of a New Cytoplasmic Male-Sterility System in Brassica juncea through Wide Hybridization

A new cytoplasmic male-sterility system was developed in an oilseed Brassica, viz. B. juncea var. ‘Pusa Bold’ with the cytoplasmic background of a wild species, Diplotaxis siifolia, obtained through wide hybridization. The synthetic alloploid (D. siifolia×B. juncea: 2n = 56, D³D³AABB) was repeatedly backcrossed to B. juncea to achieve cytoplasmic substitution. The CMS plants resembled the cultivar in growth and morphology. The flowers had narrow sepals and petals and short, shrivelled anthers which failed to dehisce. The meiotic process appeared to be normal. The microspores degenerated at an early stage after tetrad formation. Female fertility in the CMS plants was as good as in the cultivar. Female transmission of sterility confirmed it to be cytoplasmically encoded.     

Drought effects on yield and its components in Indian mustard (Brassica juncea L.)

Effects of drought on yield and yield components were investigated during the spring season 2000–2001 by growing 14 Indian mustard genotypes under irrigated and rain-fed conditions at Bharatpur and Jobner. A disease and pest management schedule was followed when required. The drought susceptibility index (DSI) for seed yield and component characteristics was calculated to characterize the relative tolerance of genotypes. Plant height, primary branches, secondary branches per plant, 1000-seed weight and seed yield were reduced under rain-fed conditions. The top five genotypes at Bharatpur that showed tolerance to moisture stress for seed yield, as indicated by their lowest DSI, were, in descending order PSR-20, PRO-97024, JMMWR-941, IS-1787 and PCR-7, whereas at Jobner these were JMMWR-941, RC-1446, PSR-20, RH-819 and 'Varuna'. Of these, PSR-20 and JMMWR-941 were among the top six at both locations. These genotypes also showed relatively low DSI for one or more characteristics, such as primary branches per plant, secondary branches per plant, harvest index and seed : husk ratio. Genotypes with the lowest DSI, particularly for seed yield at both locations, would serve as useful donors in the breeding programme for improving the drought tolerance of existing Indian mustard cultivars.     

Resynthesis of Brassica juncea through interspecific crosses between B. rapa and B. nigra

The objective of this study was to broaden the genetic base in oleiferous Brassica juncea by resynthesis, using 10 diverse parental lines of oleiferous B. rapa and two lines of B. nigra of both Indian and exotic origin. Out of 14 crosses attempted using B. rapa as the female parent, eight were successful. Embryo rescue was necessary to obtain interspecific plants. A total of 29 fertile interspecific plants were obtained after colchicine treatment. In the S₂ generation, the expression of component characters in the majority of the resynthesized plants showed a negative trend. The resynthesized B. juncea lines are being maintained through repeated selfing and selection at each generation for desirable plant types. This process will continue till the progeny lines of the desirable plants achieve uniformity.     

Introgression of Resistance to Shattering in Brassica napus from Brassica juncea through Non-Homologous Recombination

Genetic information conffering non- shattering of siliques has been introgressed in rapeseed (Brassica napus) following; interspecific hybridization between Brassica juncea and B. napus. A reconstituted B. napus plant with complete non-dehiscence of its fruits was isolated in the BC-, generation. This plant had normal meiosis and formed 19 bivalents. Its seed fertility, however, was low (23 percent). It is suggested that the gene(s) for shattering-resistance were incorporated into a B. oleracea chromosome following allosyndetic; chromosome pairing and. segmental exchange between B. nigra and B. oleracea chromosomes in the initial interspecific AABC hybrid.     

Interspecific hybridization of Brassica napus and B. juncea through ovary,ovule and embryo culture

Summary Employing in vitro culture of ovaries, ovules and embryos, interspecific hybrids have been obtained amongst two important oilseed crops, Brassica napus x B. juncea and their reciprocal. The test-tube hybrid plants have been transferred to the field, and reared to maturity. The F₁ seeds obtained from the hybrid ovaries showed normal germination, and the hybrid plants exhibited a range of variation of characters.     

Interspecific hybridization inBrassica juncea × Brassica hirta using embryo rescue

Summary Interspecific hybrids have been obtained in an otherwise incompatible cross betweenBrassica juncea × Brassica hirta through the in vitro culture of hybrid ovules and ovaries. The best response was observed from ovules and ovaries cultured 10–15 and 5–7 days after pollination respectively on a basal medium supplemented with indoleacetic acid, kinetin and casein hydrolysate. In some cases the basal cut end of the ovaries proliferated to form callus and shoots. The in vitro-derived hybrid seeds varied in their colour, size and shape, and the F₁ plants in the field showed a large diversity in their morphological traits. The hybrids were sterile, and had an intermediate number of chromosomes (2n=30).     

Phenotypic variation and the relationships among mustard (Brassica juncea L.) germplasm from Pakistan

An investigation was conducted to determine the extent of diversity and relationships among the Brassica juncea germplasm from Pakistan using morphological characters. A total of 52 accessions, including the collected germplasm as well as commercial cultivars/improved lines, were studied under field conditions at Tsukuba, Japan during 1995 and 1996. All the accessions were characterized for 35 agro-morphological characters from seedling emergence to crop maturity. Morphological data were analyzed by numerical taxonomic techniques using two complementary procedures: cluster and principal component analyses. Phenograms based on Euclidean distance placed the accessions into six groups during both years. Landrace groups were primarily associated with morphological differences among the accessions and secondarily with the breeding objectives and horticultural uses. The mustard germplasm collected from Pakistan showed a comparatively low level of phenotypic variation amongst themselves and were genetically similar to the oilseed cultivars. However, the oilseed forms and vegetable cultivars were genetically distinct. This study revealed that the evaluated germplasm appears to have a narrow genetic base which undergoes a high level of genetic erosion. This is perhaps due to the use of the same ancestors in the selection of new lines for similar horticultural traits, replacement by major crops and changes in agricultural land uses. This revised version was published online in July 2006 with corrections to the Cover Date.     

Interspecific hybridization between Brassica carinata and Brassica rapa

The crossability between Brassica carinata (BBCC, 2n=34) and Brassica rapa (AA, 2n=20), and the cytomorphology of their F₁ hybrids were studied. Hybrids between these two species were only obtained when B. carinata was used as the female parent. The hybrid plants exhibited intermediate leaf and flower morphology, and were found to be free from white rust and Alternaria blight diseases. One of the four F₁ plants was completely male sterile, while the remaining plants had 4.8, 8.6, and 10.9% stainable pollen, respectively. No seed was produced on hybrid plants under self pollination or in backcrosses; but seed was obtained from open pollination. The occurrence of the maximum of 11 bivalents as well as up to 44.8%) of cells with multivalent associations in the form of trivalents (0‐2) and a quadrivalent (0‐1) in the trigenomic triploid hybrid (ABC, 2n = 27) revealed intergenomic homoeology among the A, B and C genomes. Meiotic analysis of F₁ hybrids indicated that traits of economic importance, such as disease resistance, could be transferred from B. carinata to B. rapa through interspecific crosses.     

Glucosinolate content in interspecific crosses of Brassica carinata with B. juncea and B. napus

Brassica carinata A. Braun is a highly productive oilseed crop in the Ethiopian highlands, but the seed has a high 2-propenyl glucosinolate content, which is undesirable. The objective of this study was to introgress, through interspecific crosses, genes for low 2-propenyl glucosinolate content from the B genome of B. juncea and C genome of B. napus into the B. carinata B and C genomes and thus develop low glucosinolate B. carinata. The cross [(B. carinata×B. juncea) ×B. carinata] yielded plants that contained only ～ 20 μmoles of 2-propenyl glucosinolate, which was an 85% reduction compared with levels in B. carinata seed. Plants of the [(B. carinata×B. napus) ×B. carinata] cross had normal high concentrations of 2-propenyl glucosinolate. Backcross plants of both interspecific crosses also contained 3-butenyl and 2-hydroxy-3-butenyl glucosinolates. The results of these crosses suggested that genes for glucosinolate synthesis were located on B genome chromosomes of B. carinata because B. napus C genome introgressions did not result in reductions of total glucosinolate contents. The total alkenyl glucosinolate content of one F₃ family of the B. juncea backcross was similar to that of the B. juncea parent. It was concluded that through further selection in this family, B. carinata plants could be identified that would be basically free of 2-propenyl glucosinolate, and have a low total alkenyl glucosinolate content.     

Differential response and genes for resistance to Peronospora parasitica (downy mildew) in Brassica juncea (mustard)

The response of a wide range of Brassica juncea accessions to 14 isolates of Peronospora parasitica, 12 from India (IP00A, IP02, IP03, IP04, IP04A, IP05, IP05B, IP33 and IP33A were derived from B. juncea; IP09, IP14 and IP13A from B. rapa) and two from B. napus in the UK (R1 and P003), was screened. Sixteen differential host response groups to these isolates (classified as groups A‐P) were identified. Groups‘A’and‘B’expressed the widest resistance profiles to these isolates. Group‘A’was susceptible to isolates IP05 and IP05B, moderately resistant to isolate IP33 and resistant to all other isolates. Group‘B’was susceptible to isolates IP03, IP04 and IP04A, and resistant to the other isolates. Putative homozygous lines resistant to all 14 isolates were selected from the F₄ progeny of crosses involving lines RESBJ‐200 from group‘A’(selection from cv. Kranti) and RESBJ‐190 from group‘B’(selection from cv. Krishna). Both selections were selfed and tested for uniformity of reactions to all isolates for three generations. The resistance of RESBJ‐200 to isolates IP00A, IP04A and IP33A seems to be conditioned by single dominant genes. The resistance of RESBJ‐190 to isolates IP00A, IP05B and IP33A was also conditioned by single dominant genes. The gene for resistance to IP00A and IP33A in RESBJ‐200 seems to be independent of the genes for resistance to the same isolates in RESBJ‐190. The new genes for differential resistance to P. parasitica will be of value in future studies of the genetics of the host‐pathogen interaction and for breeding for disease resistance.     

Genetic diversity in mustard (Brassica juncea L.) germplasm from Pakistan as determined by RAPDs

The genetic diversity and the relationships among a collection of mustard (B. juncea) germplasm, including 41 accessions collected from Pakistan, 6 oilseed cultivars/ lines and 5 Japanese vegetable cultivars, were evaluated using random amplified polymorphic DNA (RAPD) markers. A total of 198 polymorphic amplified products were obtained from 30 decamer primers. Of these, 14 were unique to the accession PAK-85835 and 37 were specific to PAK-85839. Based on pair-wise comparisons of RAPD amplification products, genetic similarity was estimated using similarity coefficients of Nei & Li (1979) and a dendrogram was constructed using the unweighted pair-group method with arithmetic averages (UPGMA). Cluster analysis based on these genetic similarities placed most of the collected germplasm and oilseed cultivars/lines close to each other, showing a low level of polymorphism between the oilseed accessions collected in Pakistan. However, the clusters formed by the oilseed collections and cultivars were distinct from those formed by the vegetable cultivars. A low level of genetic variability of oilseed mustard in Pakistan was attributed to the selection for similar traits and horticultural uses. The farmers' preference for more remunerative crops and perhaps the close parentage of these accessions further contributed towards their little diversity. The study demonstrated that the RAPD is a simple and fast technique to compare the genetic relationships and the patterns of variation among accessions of this crop. This revised version was published online in August 2006 with corrections to the Cover Date.     

Transfer of powdery mildew resistance from Brassica carinata to Brassica oleracea through embryo rescue

Interspecific hybrid plants and backcross 1 (BC₁) progeny were produced through sexual crosses and embryo rescue between Brassica carinata accession PI 360883 and B. oleracea cvs Titleist’and‘Cecile’to transfer resistance to powdery mildew to B. oleracea. Four interspecific hybrids were obtained through application of embryo rescue from crosses with B. carinata as the maternal parent, and their interspecific nature confirmed through plant morphology and random amplified polymorphic DNA (RAPD) analysis. Twenty‐one BC₁ plants were obtained through sexual crosses and embryo rescue although embryo rescue was not necessary to produce first backcross generation plants between interspecific hybrids and B. oleracea. All interspecific hybrids and eight of the BC₁ plants were resistant to powdery mildew.     

Development of Yellow Seeded Brassica napus Through Interspecific Crosses

Yellow seeded Brassica napus was developed through interspecific crosses with the two mustard species, B. juncea and B. carinata. The objective of these two interspecific crosses was the introgression of genes for yellow seed colour from the A genome of B. juncea and C genome of B. carinata into the A and C genomes of B. napus, respectively. The interspecific F₁ generations were backcrossed to B. napus in an attempt to eliminate B genome chromosomes and to improve fertility. Backcross F₂ plants of the (B. napus×B. juncea) ×B. napus cross were then crossed with backcross F₂ plants of the (B. napus×B. carinata) ×B. napus cross. The objective of this intercrossing was to combine the A and C genome yellow seeded characteristics of the two backcross populations into one genotype. The F₂ generation of the backcross F₂ intercrosses was grown in the field, plants were individually harvested and visually rated for seed colour. Ninety-one yellow seeded plants were identified among the 4858 plants inspected. This result indicated that the interspecific crossing scheme was successful in developing yellow seeded B. napus.     

Bud pollination and hybrid seed production in detergent-induced male sterile plants of Brassica juncea

The efficacy of a synthetic detergent (Surf Excel) as a potential chemical hybridizing agent in Brassica juncea was studied. Foliar sprays with various concentrations of the detergent caused reductions in plant height, number of branches and leaves per plant, size of leaves, anther size, pollen per flower, ovules per flower, pollen fertility, fruits per plant, fruit size, seeds per fruit, total yield per plant and 100 seed weight as compared with those of untreated plants. The style in the floral buds of plants sprayed with different concentrations of Surf Excel elongated and so the receptive stigma protruded from the buds which facilitated cross‐pollination by honey bees. The plants sprayed once with 2% Surf Excel exhibited an elongated style with a raised receptive stigma and 100% pollen sterility without causing a significant reduction in total yield.     

Hybridization in Brassica juncea × Brassica campestris through ovary culture

Summary Ovary culture has been employed for the production of interspecific hybrids of a partially compatible cross of Brassica juncea (2n=36) × Brassica campestris (2n=20). Five to seven days old ovaries cultured on White's medium supplemented with casein hydrolysate (300 mg/l) and sucrose (5%) produced more seeds than any other media tried, but seed development was better on media fortified with plant hormones. The seed yield was better in B. juncea × B. campestris than their reciprocal cross. The plants obtained from ovary-derived seeds were transferred to the field; they were intermediate in some morphological characters and chromosome number (2n=28) as compared to their parents. The flower buds generally did not open and had poorly developed anthers with mostly sterile pollen. The pod size/setting was very much reduced, but healthy seeds were obtained.