Linked epistasis for six quantitative traits in Indian mustard (Brassica juncea (L.) Czern & Coss)

Summary Gene effects, and interactions, and associations between days-to-flower initiation and maturity, number of secondary branches and siliquae per plant, and 1,000-seed weight and yield per plant were studied in a cross of Indian mustard (Brassica juncea (L.) Czern & Coss) using the parents and F₁, F₂, F₃, B₁, B₂, B₁₁, B₁₂, B₂₁, B₂₂, B_1S, B_2S, B₁F₁, B₂F₁, B_1bip, B_2bip, F₂P₁, F₂F₁, and F_2bip generations. A linked digenic model was adequate for all characters studied. According to this model, the main effects, additive and interactions between linked pairs of genes, were present in varying proportions for days-to-flower initiation and maturity and number of siliquae per plant. The contribution of linked epistatic effects, however, was much greater than that of additive effects. Dominance effects contributed significantly to the inheritance of days-to-flower initiation. Duplicate epistasis was observed for all traits except 1,000-seed weight where epistasis was of the complementary type. A complete association among the genes of similar effect (increasing or decreasing) was observed for number of secondary branches and siliquae, and yield per plant. Coupling phase linkage was observed for days-to-flower initiation whereas repulsion phase linkage was observed for daysto-maturity and 1,000-seed weight.     

A RFLP-based linkage map of mustard [Brassica juncea (L.) Czern. and Coss.]

 A genetic linkage map of Brassica juncea was constructed based on restriction fragment length polymorphism (RFLP) detected by anonymous cDNA markers from B. napus, using a segregating F₁-derived doubled haploid (DH) progeny from a cross between a canola-quality mustard line (J90-4317) and a high-oil-content mustard line (J90-2733). The RFLP probes consisted of 229 cDNA probes from B. napus and a B. napus tandem repeat sequence, RDA2. The map consisted of 343 marker loci arranged in 18 major linkage groups plus five small segments with two to five marker loci, covering a total map distance of 2073 cM. Twenty-four percent of the markers were dominant in nature. Sixty-two percent of the marker loci were duplicated, and the majority were involved in inter-linkage group duplications, illustrating that complex duplications and subsequent rearrangements occurred after allopolyploidy. Deviation from the Mendelian segregation ratio for a DH population was observed for 27% of the markers. Two-thirds of these markers with a skewed segregation were clustered in 6 linkage groups and two unassigned segments. The overall average marker interval of the B. juncea map reported here was 6.6 cM, which would provide a marker density satisfactory for efficient use of the map in breeding applications, such as tagging of important agronomic traits and marker-assisted selection. Received: 14 May 1996 / Accepted: 11 October 1996     

Effect of nitrogen and chlormequat chloride on the seed yield and oil content of mustard (Brassica juncea L. Czern & Coss)

The growth of mustard was increased significantly when treated with up to 80 kg N ha^–1 (N₈₀). Spraying with (2-chloroethyl) trimethylammoniumchloride (chlormequat chloride) increased seed yield and seed protein content. Spraying nitrogen fertilized plots with chlormequat chloride, increased leaf area, leaf area ratio, leaf area duration, number of siliquae plant^–1, number of seeds siliqua^–1 and length of siliqua. Reducing, non-reducing and total sugars in the leaves at 80 days after sowing were also affected significantly. Chlorophyll a, b and total chlorophyll were little affected. The number of siliquae plant^–1 was highly correlated with seed yield in both the seasons of experimentation. The correlation coefficient value () was 0.586 in 1982/83 and 0.912 in 1983/84.The total accumulation of nutrients, i.e. nitrogen, phosphorus and potassium in seed and straw was significantly affected by N₈₀ × chlormequat chloride interaction.     

Generation mean analyses for flowering and maturity in Indian mustard (Brassica junces (L.) Czern & Coss)

Summary Components of generation means were partitioned for days to flower initiation and maturity in three crosses of Indian mustard (Brassica juncea (L.) Czern and Coss) cultivars. A linked digenics model was adequate for flowering in cross II and maturity in Cross I. All three types of digenic interactions among the linked pairs of genes, additive X additive (i), additive X dominance (j) and dominance X dominance (1), contributed significantly in the inheritance of flowering in cross II and maturity in cross I. A complete association among the genes of greater effects in higher mean parent was detected for flowering in cross II and maturity in Cross I. Duplicate epistasis was evident for flowering in Crosses I and II and maturity in Crosses I and III.Inadequacy of all the fitted models for days to flowering in Cross III and maturity in Cross II indicated the presence of higher order interactions.Part of PhD (plant breeding) Thesis, submitted by senior author to GBPUAT, Pantnagar (Nainital) U.P., India (unpublished). Research paper No. 4262     

Alternaria incidence in some alloplasmic lines of Indian mustard (Brassica juncea (L.) Coss.)

Summary The cytoplasmic substitution lines of Brassica juncea (L.) Coss were evaluated for their field resistance to Alternaria blight (Alternaria brassicae). The euplasmic B. juncea cv. RLM 198 had a mesothetic reaction while alloplasmic B. juncea lines with cytoplasms of B. campestris, B. chinensis, and B. japonica were highly susceptible. B. nigra cytoplasm did not have any effect on the disease reaction of the B. juncea genome. However, the alloplasmic lines with the cytoplasm of B. napus and B. carinata revealed a comparatively higher degree of resistance. The study underlined the utility of cytoplasmic manipulations in modifying the phenotypic expression of nuclear genes.     

Influence of paclobutrazol on growth and development of fruit in Brassica juncea (L.) Czern and Coss

Foliar spraying of paclobutrazol(5, 10 and 20 g mL^–1) on Brassica juncea (cv. RLM 514) plants, at the green floral bud stage of the terminal inflorescence, exerted a profound effect on growth and metabolism of fruits (siliquae). Paclobutrazol caused a decrease in siliqua length but enhanced significantly siliqua width, seed size (diameter) and seed number per siliqua. Paclobutrazol increased dry matter production and also hastened the relative growth rate of the siliqua wall and seed. In paclobutrazol-treated plants the siliquae were darker green, and during active phases of growth both siliqua wall and seed exhibited higher total chlorophyll (Chl) content, decreased chl a/b ratios, greater hill reaction activity of chloroplasts and increased activities of -amylase, -amylase and invertase than controls. Paclobutrazol enhanced the levels of total soluble sugars, starch, protein and free amino acids in mature seeds. The seed oil content increased by 2 to 3 per cent with a treatment of 10 g mL^–1 pacolobutrazol.     

Mapping of QTLs for oil content and fatty acid composition in Indian mustard [Brassica juncea (L.) Czern. and Coss.]

An AFLP linkage map of Brassica juncea (L.) Czern and Coss was constructed using 88 recombinant inbred lines (RILs) from a cross between an Indian cultivar ‘Varuna’ and an accession from Poland ‘BEC-144’. The map included 91 AFLP markers organized on 19 linkage groups covering a total map distance of 1679.1 cM. A total of 14 QTLs were detected for oil content (2 QTLs), erucic acid (2 QTLs), eicosenoic acid (2 QTLs), linolenic acid (3 QTLs), linoleic acid (3 QTLs) and palmitic acid (2 QTLs). A specific genomic region on LG2 was associated with contents of three fatty acids: erucic acid, eicosenoic acid and linoleic acid. Some of the markers showed absolute linkage with the QTLs associated with the levels of linolenic acid, linoleic acid and oil content. These markers may be used for improvement of fatty acid profile of B. juncea.     

Molecular cloning and characterization of five annexin genes from Indian mustard (Brassica juncea L. Czern and Coss)

Plant annexins constitute a multigene family having suggested roles in a variety of cellular processes including stress responses. We have isolated and characterized five different cDNAs of mustard, Brassica juncea (AnnBj1, AnnBj2, AnnBj3, AnnBj6 and AnnBj7) encoding annexin proteins using a RT-PCR/RACE-PCR based strategy. The predicted molecular masses of these annexins are ～36.0 kDa with acidic pIs. At the amino acid level, they share high sequence similarity with each other and with annexins from higher plants. Phylogenetic analysis revealed their evolutionary relationship with corresponding orthologous sequences in Arabidopsis and deduced proteins in various plant species. Expression analysis by semi-quantitative RT-PCR revealed that these genes are differentially expressed in various tissues. The expression patterns of these genes also showed regulation by various stress conditions such as exposure to signaling molecules, salinity and oxidative stress and wounding. Additionally, the in silico promoter analysis (of AnnBj1, AnnBj2 and AnnBj3) showed the presence of different cis-responsive elements that could respond to various stress conditions. These results indicate that AnnBj genes may play important roles in adaptation of plants to various environmental stresses.     

C-genome chromosome substitution lines in Brassica juncea (L.) Coss

Spontaneously developed C genome substitution lines of Brassica juncea (L.) Coss were identified in the F₇ generation of an interspecific cross between B. juncea (AABB; 2n=36) and B. napus (AACC; 2n=38). These lines were meiotically stable (2n=36) and had normal male and female fertility. They were characterized by certain specific traits like bolting habit, leaf aberrations and variation for erucic acid content. Cytological evidence indicated that at least three lines has substitution for different chromosomes. The substitution stocks characterized in this study are important for their use in practical crop breeding and certain basic genetic studies.     

Isolation and characterisation of satellite DNA from Brassica juncea (L.) Czern

Nuclear DNA isolated from hypocotyls (H), proliferating callus (PC) and differentiating callus (DC) of Brassica juncea contains a satellite DNA which can be resolved in actinomycin-D/CsCl gradients. The satellite DNA undergoes changes, when an in vitro culture is raised from hypocotyl tissue and forms a higher percentage of the genome in PC and DC than in mature differentiated tissue (hypocotyl). All the three satellite DNAs are GC-rich compared to main band DNAs. Satellite DNA of H has higher Tm and GC content than that of the PC and DC satellites. A 200 bp basic repeat unit from hypocotyl nuclear DNA has been cloned and characterised.     

Calcium supplementation modulates arsenic-induced alterations and augments arsenic accumulation in callus cultures of Indian mustard (Brassica juncea (L.) Czern.)

In the present study, the effect of arsenate (AsV) exposure either alone or in combination with calcium (Ca) was investigated in callus cultures of Brassica juncea (L.) Czern. cv. Pusa Bold grown for a period up to 24 h. The AsV?(250 μM) + Ca (10 mM) treatment resulted in a significantly higher level of As (464 μg g^?1 dry weight (DW)) than AsV without Ca (167 μg g^?1 DW) treatment at 24 h. Furthermore, AsV + Ca-treated calli had a higher percent of AsIII (24–47%) than calli subjected to AsV treatment (12–14%). Despite this, AsV + Ca-treated calli did not show any signs of hydrogen peroxide (H₂O₂) accumulation or cell death upon in vivo staining, while AsV-exposed calli had increased H₂O₂, shrinkage of cytoplasmic contents, and cell death. Thus, AsV treatment induced oxidative stress, which in turn elicited a response of antioxidant enzymes and metabolites as compared with control and AsV + Ca treatment. The positive effects of Ca supplementation were also correlated to an increase in thiolic constituents', viz., cysteine, reduced glutathione, and glutathione reductase in AsV + Ca than in AsV treatment. An analysis of selected signaling related genes, e.g., mitogen-activated protein kinases (MAPK3 and MAPK6) and jasmonate ZIM-domain (JAZ3) suggested that AsV and AsV + Ca followed variable pathways to sense and signal the As stress. In AsV-alone treatment, jasmonate signaling was seemingly activated, while MAPK3 was not involved. In contrast, AsV + Ca treatment appeared to specifically inhibit jasmonate signaling and activate MAPK3. In conclusion, Ca supplementation may hold promise for achieving increased As accumulation in plants without compromising their tolerance.     

Regeneration of plantlets from mesophyll protoplasts of Brassica juncea (L.) Czern

Isolated mesophyll protoplasts of Brassica juncea (L.) Czern., cv. RLM 514 upon culture in suitable growth medium, regenerated cell wall, underwent cell division and formed cellular colonies. Subsequent induction of embryoid (embryogenesis) and shoot bud (organogenesis) formations in such cell masses resulted in regeneration of 186 and 42 plantlets respectively.Abbreviations NT Nagata and Takebe, 1971 - B5 Gamborg et al. 1968 - KM Kao and Michayluk, 1975 - GK₂ Schenck and Hoffmann, 1978 - MS Murashige and Skoog, 1962 - BAP 6-benzyladenine - 2, 4-D 2, 4-dichlorophenoxyacetic acid - NAA napthaleneacetic acid - GA₃ gibberellic acid     

Alleviation of Cadmium Toxicity in Brassica juncea L. (Czern. & Coss.) by Calcium Application Involves Various Physiological and Biochemical Strategies

Calcium (Ca) plays important role in plant development and response to various environmental stresses. However, its involvement in mitigation of heavy metal stress in plants remains elusive. In this study, we examined the effect of Ca (50 mM) in controlling cadmium (Cd) uptake in mustard (Brassica juncea L.) plants exposed to toxic levels of Cd (200 mg L⁻¹ and 300 mg L⁻¹). The Cd treatment showed substantial decrease in plant height, root length, dry weight, pigments and protein content. Application of Ca improved the growth and biomass yield of the Cd-stressed mustard seedlings. More importantly, the oil content of mustard seeds of Cd-stressed plants was also enhanced with Ca treatment. Proline was significantly increased in mustard plants under Cd stress, and exogenously sprayed Ca was found to have a positive impact on proline content in Cd-stressed plants. Different concentrations of Cd increased lipid peroxidation but the application of Ca minimized it to appreciable level in Cd-treated plants. Excessive Cd treatment enhanced the activities of antioxidant enzymes superoxide dismutase, ascorbate peroxidase and glutathione reductase, which were further enhanced by the addition of Ca. Additionally, Cd stress caused reduced uptake of essential elements and increased Cd accumulation in roots and shoots. However, application of Ca enhanced the concentration of essential elements and decreased Cd accumulation in Cd-stressed plants. Our results indicated that application of Ca enables mustard plant to withstand the deleterious effect of Cd, resulting in improved growth and seed quality of mustard plants.     

A hypocotyl-derived somatic embryogenic system in Brassica juncea Czern & Coss and its manipulation for enhanced storage lipid accumulation

A simple and reproducible protocol for induction, growth and development of somatic embryos from hypocotyl explants of Indian mustard (Brassica juncea L. Czern & Coss) var. RLM 198 is reported. The HDSE (Hypocotyl-derived somatic embryos) were fleshy globular to torpedo structures that were maintained by regular subculturing every three weeks. These embryos developed non-synchronously into the heart shaped-stage while some were matured into a green cotyledon-stage bearing embryos in the same medium. The HDSE accumulated as much as 50.2% lipid content on a dry weight basis at 14 DAC (days after culture) using a culture medium supplemented with 10% PEG (Polyethylene glycol 6000) in comparison to less than 15% lipid content in 2% sucrose (control) or 20 μM ABA (abscisic acid). An increase in total soluble sugar content was observed with 2.5% PEG and increasing PEG concentration caused a decrease in their contents in HDSE. The activities of invertase, acetyl CoA carboxylase and 1-¹⁴C-acetate incorporation into lipids in HDSE were enhanced significantly in the culture medium containing 10% PEG. The content of triacylglycerols in HDSE was maximum with 10% PEG supplemented culture medium. The wax content in HDSE increased progressively with an increase of PEG concentration in the culture medium. The ABA and PEG supplementation increased the content of membrane lipids when the data was expressed on a 100 g dry weight basis. The proportion of palmitate and erucate decreased and that of oleate, linoleate and linolenate increased at 14 DAC in HDSE in 10% PEG supplemented culture medium. Thus, the manipulation of culture conditions significantly altered total lipid content, membrane lipid composition and the quality of storage lipids. This revised version was published online in August 2006 with corrections to the Cover Date.     

芥菜类作物的遗传多样性

芥菜类是我国重要的经济作物,遗传资源十分丰富。现就目前国内外芥菜类作物遗传多样性研究的现状进行综述,并着重阐述了芥菜类作物的分类、分布以及遗传多样性研究方法,如形态学分析法、同工酶分析法、RAPD法和RFLP法。同时对芥菜类作物遗传多样性研究存在的问题和今后的研究方向进行了探讨。     

Morphological and physio-biochemical characterization of Brassica juncea L. Czern. & Coss. genotypes under salt stress

Abstract

Soil salinity is one of the major factors responsible for the low productivity of crop plants and has become an increasing threat for agriculture. In this context, the selection of tolerant genotype/s may be one of the remedies. Keeping this in view, the effect of NaCl (0–120 mM) stress on shoot length (SL) plant^?1, area (A) leaf^?1, leaf area index (LAI), fresh weight (FW) and dry weight (DW) plant^?1, stomatal conductance (g_s), net photosynthetic rate (P _N), total chlorophyll (Chl) content, malondialdehyde (MDA) content, sensitivity rate index (SRI), leaf- nitrogen (N), potassium (K) and sodium (Na) content, leaf-K/Na ratio, nitrate reductase (NR: EC.1.6.6.1) and ATP-sulphurylase (ATP-S: EC.2.7.7.4) activities and proline (Pro) and glycinebetaine (GB) content of ten genotypes of Brassica juncea L. was studied at 55 and 65 days after sowing (DAS). NaCl treatments decreased all the above parameters, except Pro, GB, MDA, Na and SRI at both stages. Salt stress resulted in accumulation of Pro and GB, in all genotypes. The magnitude of increase in both osmolytes (Pro and GB) was higher in genotype G₈ than the other genotypes. Salt stress increased MDA and Na content while it decreased Chl, N and K content and K/Na ratio, Chl content, NR and ATP-S activities in all genotypes. But the magnitude of increase in MDA and Na content and decrease in SL plant^?1, A leaf^?1, LAI, P _N, g_s, Chl content and NR and ATP-S activities in genotype G₈ was more than that of other genotypes. These results suggest that the salt-tolerant genotype may have better osmotic adjustment and protection from free radicals by increasing the accumulation of Pro and GB content with overproduction of N and K and higher K/Na, NR and ATP-S activities under salinity stress.