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.     

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     

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.     

Role of Ca2+ on Growth of Brassica campestris L. and B.juncea (L.) Czern & Coss under Na+ Stress

Root and shoot growth of Brassica campestris L.and B.juncea increased significantly(P0.01) with enhanced Ca2+ treatment along with 60 mM NaCl in the root medium.The maximum fresh mass of shoot and root in B.juncea was recorded at 10 mM Ca2+ concentration.The relative growth rate of shoot of both species reached its maximum at 8 mM of Ca2+ concentration.Average rate of Ca2+ intake(Ca) was higher in B.juncea than B.campestris.In B.juncea,the average transport of Ca2+ to shoot increased by 19%,38%,119%,125% and 169% compared with the control.Furthermore specific utilization rate of Ca2+ was higher in B.juncea than B.campestris.In B.campestris it increased by 9%,32%,41% and 59% at 4,6,8,and 10 mM of calcium in comparison to 2 mM Ca2+ treatment.At 4,6,8 and 10 mM of Ca2+ application,the increase in the leaf area ratio was 10,17,23 and 30%,respectively.In the shoot and root portions of B.campestris and B.juncea,Ca2+ had a linear relationship with potassium and sulfur,whereas it was in antagonism with sodium ion.     

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.     

A Study on the Volatile Flavor Constituents of Sichuan Preserved Vegetable (Brassica juncea Czern et Coss.)

The volatile constituents of Sichuan Preserved vegetable (Brassica juncea) were separated by a distillation-extraction device, and 41 compounds were identified by tile GC-MS-DS, and with the retention index of GC. The samples were obtained by N2 blast adsorption with the Porapak Q as adsorbent. They were then eluted with ether and separated by the column chromatography of silica gel. The concentrate fractions obtained were again eluted with benzene and ether. It was identified that the con- centrate fractions possessed the characteristic odor of Preserved vegetable (Brassica juncea), and that the characteristic odor compounds were: 3-butenonitrile, phenyla- cetonitrile, 3-phenypropiononitrile, allyl isothiocyanate, 2-phenylethyl isothiocyanate, di- methyl trisulfide, 1-allyl-4-methoxybenzene, 1-methoxy-4-(1-propenyl)-benzene, 7-methoxybenzofuran. The result of this study may be helpful to improve the aroma quality of preserved vegetable, to develop its new varieties, and to research the aroma of food stuffs.     

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.     

Genetic architecture of yield and components of yield in mustard (Brassica juncea (L.) Czern & Coss.)

Summary The relative ability of cross- and self-pollen to achieve fertilisation in Brassica was studied by making double pollinations using cross-pollen carrying a dominant seedling marker gene. With simultaneous self- and crosspollination 12–40% self-seed was set, but when cross-pollen was applied to the stigma four hours before self-pollen, only 2–4% self-seed was obtained. In two plants to which cross-pollen was applied at various time intervals after self-pollen there was a tendency for the percentage of self-seed to increase as the time interval increased. In a third plant this trend was not apparent, probably because of a greater degree of self-incompatibility. The competitive advantage of the first pollen to arrive on the stigma is discussed in relation to the strength of the self-incompatibility and the sib problem in F₁ hybrid brassicas.     

影响茎用芥菜愈伤组织诱导和植株再生的因素

茎用芥菜子叶培养在MS 0.25 mg·L-1NAA 0.5 mg·L-16-BA 0.25 mg·L-12,4-D培养基上,可获得较高质量的愈伤组织.愈伤组织培养在MS 1～1 mg·L-16-BA 0.2 mg·L-1NAA培养基上分化频率为8.3%,而在加有羧苄青霉素和头孢霉素的培养基上,最高分化频率可达52.4%.将获得的再生植株转移到1/2MS 0.1 mg·L-1 NAA的生根培养基中,可获得完整植株.     

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.     

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.     

Proline status of genetically stable salt-tolerant Brassica juncea L. somaclones and their parent cv. Prakash

Three Brassica juncea L. somaclones (SR-1, -2 and -3) selected in vitro for NaCl-tolerance, non-selected somacone (CP-5) and parent cv. Prakash were characterized for their free proline contents in the absence of stress and as a function of increasing salt stress. In the R₀ generation, SR-3 somaclone had ca. three times higher free proline as compared to parent Prakash and other somaclones. Somaclone, SR-1, turned out to be sterile. The other somaclones were carried forward to the R₂ generation after making selections for yield and yield components in the R₁ generation. SR-3 bred true for its high proline accumulating characteristic. The somaclone SR-3 thus had a stable genetic variation for proline overproduction. Free proline content in 7-day-old whole seedlings and 6-week-old plant leaf tissue, increased with the increase in salt stress in all the lines but at differential rates. The magnitude of increase in free proline was much higher in SR-3 lines as compared to parent Prakash and SR-2 salt-tolerant somaclones. Under salt stress, in leaf tissue, one of the SR-3 derived lines (SR3P6-2) accumulated as much as 269 moles of free proline as compared to ca. 20 moles per g dry weight in parent Prakash and SR-2 line. It was interesting to note that there was a critical point concentration of NaCl beyond which the endogenous level of free proline rose sharply. Somaclonal lines (SR3P6-2, SR2P1-2 and CP5-2) which were found to have higher salt-tolerance indices, also had higher critical points as compared to the other relatively salt sensitive genotypes. The relationship between relative water content and osmotic potential of leaves under saltstress also showed a relatively higher degree of osmotic adjustment in the selected somaclones, the maximum being in SR-3 derived lines.     

Study of factors affecting the culture of Brassica napus L. and B. juncea Coss. mesophyll protoplasts

Various factors affecting the culture of Brassica napus and B. juncea mesophyll protoplasts were examined in order to develop suitable culture media for these species. The basic components (salts and vitamins) of culture media K3 and Kao best supported cell division and colony development in protoplast culture of both species. The addition of casamino acids to Kao's medium resulted in colony browning in B. napus genotypes. B. napus protoplasts grew well with glucose as the osmotic stabilizer, whereas B. juncea protoplasts responded better to sucrose. High NAA and low 2,4-D combinations were effective in stimulating colony growth. Colony development was rapid for a range of genotypes cultured with these recommendations in these media and plant regeneration was obtained from protoplast-derived calli in both species.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - NAA -naphthaleneacetic acid - BAP 6-Benzylaminopurine - MES 2(N-Morpholino)ethane sulfonic acid Contribution No. 931.     

Zinc application mitigates the adverse effects of NaCl stress on mustard [Brassica juncea (L.) Czern & Coss] through modulating compatible organic solutes,antioxidant enzymes,and flavonoid content

This study examined the protective effect of Zn on salt-stressed Brassica juncea plants using some key morphological and biochemical attributes at different developmental stages (30, 60, and 90 days after treatment [DAT]). Salt stress (200?mM) caused suppression in plant height, root length, and dry weight by 58.35%, 41.15%, and 53.33%, respectively, at 90 DAT, but Zn application improved these variables by 15.52%, 16.59%, and 11.45%, respectively. Furthermore, 200?mM NaCl decreased total chlorophyll by 45.32% and relative water content by 27.62% at 90 DAT, whereas Zn application compensated the decrease in the levels of both variables. NaCl (200?mM) increased H₂O₂, malondialdehyde, and electrolyte leakage by 70.48%, 35.25%, and 68.39%, respectively, at 90 DAT, but Zn supplementation appreciably reduced these variables. Except for catalase, enzymatic antioxidant activity increased under NaCl stress. Zn application with salt further increased the activities of superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase, and glutathione-S-transferase by 33.51%, 9.21%, 10.98%, 17.46%, and 12.87%, respectively, at 90 DAT. At 90 DAT, salt stress increased flavonoids by 24.88%, and Zn supply by a further 7.68%. Overall, Zn mitigated the adverse effects of salt stress through osmotic adjustment, as well as by modulating the oxidative defense system and flavonoid contents.     

Effect of Challenge Temperature and Solute Type on Heat Tolerance of Salmonella Serovars at Low Water Activity

Salmonella spp. are reported to have an increased heat tolerance at low water activity (a_w; measured by relative vapor pressure [rvp]), achieved either by drying or by incorporating solutes. Much of the published data, however, cover only a narrow treatment range and have been analyzed by assuming first-order death kinetics. In this study, the death of Salmonella enterica serovar Typhimurium DT104 when exposed to 54 combinations of temperature (55 to 80°C) and a_w (rvp 0.65 to 0.90, reduced using glucose-fructose) was investigated. The Weibull model (LogS = −btⁿ) was used to describe microbial inactivation, and surface response models were developed to predict death rates for serovar Typhimurium at all points within the design surface. The models were evaluated with data generated by using six different Salmonella strains in place of serovar Typhimurium DT104 strain 30, two different solutes in place of glucose-fructose to reduce a_w, or six low-a_w foods artificially contaminated with Salmonella in place of the sugar broths. The data demonstrate that, at temperatures of ≥70°C, Salmonella cells at low a_w were more heat tolerant than those at a higher a_w but below 65°C the reverse was true. The same patterns were generated when sucrose (rvp 0.80 compared with 0.90) or NaCl (0.75 compared with 0.90) was used to reduce a_w, but the extent of the protection afforded varied with solute type. The predictions of thermal death rates in the low-a_w foods were usually fail-safe, but the few exceptions highlight the importance of validating models with specific foods that may have additional factors affecting survival.