Plant Regeneration from Protoplasts of Brassica Juncea L.

Protoplasts isolated enzymatically from epicotyl and growing tip of Bressica juncea divide to form callus on Kp8 medium. Plant regeneration is obtained from protoplast- derived callus of on MSD3 medium. High concentration of inositol in differentiation medium stimulates plant or shoot regeneration from the epicoty protoplast origin.     

Effects of Crude and Bioremediated Thermal Power Plant Effluents in Brassica Juncea

Crude phenol and cyanides are the major components of the effluents discharged by industries involved in the manufacture of many synthetic inorganic and organic compounds, pharmaceuticals, electroplating units, and thermal power plants.

In this study, an effort was made to use effluents as manure. The effect of various amendments (10, 40, 70, and 100%) of bioremediated and crude effluents was checked on plants of Brassica juncea. Roots, shoot lengths, and the mean dry weights were maximum in the plants irrigated with 10% treated and 10% crude effluent as compared to the control plants. Total plant phenolics increased with an increase in the amendment of the effluents in the plants irrigated with bioremediated as well as crude effluents. Applications of lower amendments of treated and untreated effluents in Brassica juncea improved the chlorophyll levels in the experimental plants and the values obtained were similar to the control plants. Effect of bioremediated and crude effluents on antioxidant enzymes of Brassica juncea showed a differential effect. The activity of antioxidant enzymes peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD) was observed to increase with an increase in the effluent concentration.

Here we demonstrate that the industrial effluents, when bioremediated of their harmful components, can also serve as a nourishment for plants, which can further help in the rehabilitation of wastelands.     

脯氨酸对不结球白菜（Brassica chinensis）镉积累的影响

采用溶液培养的方法,研究了过量Cd对不结球白菜（Brassica chinensis）幼苗内源脯氨酸含量的影响及外源脯氨酸在不结球白菜Cd积累中的作用。100μmol？L-1 Cd处理明显增加不结球白菜叶片脯氨酸（Pro）的含量,并且随着Cd胁迫时间的延长,其含量也显著上升。100μmol？L-1 Cd处理明显降低脯氨酸脱氢酶（PDH）活性,而增加δ1-吡咯啉-5-羧酸合成酶（P5CS）的活性。1~5 mmol？L-1外源Pro处理显著降低不结球白菜幼苗地上部Cd含量,对根系Cd含量无显著影响,但外源Pro处理并不能缓解过量Cd对不结球白菜幼苗生物量的抑制作用。外源Pro处理下,不结球白菜叶片中Pro含量与其地上部干重呈显著的负相关关系。     

Response to nickel in the proteome of the metal accumulator plant Brassica juncea

To gain a comprehensive understanding of the molecular mechanism of heavy metal accumulation in Brassica juncea, comparative proteomic approaches were used to analysis protein profiles in leaf tissues of 6-week-old B. juncea after exposure to 100 µM Ni. Proteomic analysis revealed that 61 protein spots showed 1.5-fold change in protein abundance after Ni exposure as compared to that of corresponding spots in control. Out of the 61 differentially expressed protein spots, 37 protein spots were ambiguously identified by matrix assisted laser desorption ionization/time of flight mass spectrometry (MALDI-TOF MS). The majority of these identified proteins were found to be involved in sulphur metabolism, protection against oxidative stress, clearly indicated that heavy metal sequestration and antioxidant system were activated by Ni treatment. The induced expression of photosynthesis and ATP generation-related proteins were also observed in plants exposed to metals, suggesting the tolerance and accumulation is an energy-demanding process. The identification of these proteins in response to Ni can lead a deep understanding of heavy metal accumulation and tolerance in B. juncea.     

印度芥菜-苜蓿间作对镉胁迫的生态响应

采用温室盆栽试验,研究了重金属镉（Cd）胁迫对印度芥菜和苜蓿间作下的土壤-植物系统的影响,同时对苜蓿的Cd饲用安全进行了评估.结果表明：在土壤Cd含量为0.37～20.37 mg·kg^-1范围内,与单作印度芥菜和苜蓿相比,间作使印度芥菜生物量降低了0.4%～11.8%,而使苜蓿生物量提高了55.3%~70.0%.土壤有效Cd主要受土壤全量Cd和种植植物种类的影响,种植方式对其影响不大.在土壤Cd含量为5.37 mg·kg^-1时,间作印度芥菜地上部Cd含量较单作提高了14.5%,而间作苜蓿地上部Cd含量较单作降低了57.1%;此时,单作和间作苜蓿地上部Cd含量分别为0.21和0.09 mg·kg^-1,均未超过饲料卫生限定标准(0.5 mg·kg^-1).在土壤Cd含量为10.37～20.37 mg·kg^-1范围内,虽然单作和间作苜蓿Cd含量均超过饲料卫生限定标准,但间作种植方式仍然使苜蓿地上部Cd含量较单作降低了2.8%～48.3%,印度芥菜地上部Cd含量也较单作降低了1.1%～48.6%.不论单作还是间作印度芥菜的Cd转运系数都远高于苜蓿.     

不同品种菜心对镉抗性的研究

以对镉（Cd)抗性不同的两个菜心品种特青60天和迟心2号为试验材料,用盆栽土培试验的方法研究了在不同浓度Cd处理下（0mg/kg,1.0mg/kg,3.0mg/kg,10.0mg/kg)Cd在菜心体内及迁移的差异,并探讨了造成差异的机理。研究结果表明：Cd处理下,特有60天茎叶中积累的Cd比迟心2号的少,在高Cd浓度（10.0mg/kg)下,与对照相比,特青60天的干重明显增加,而迟心2号的干重则显著降低。进一步分析研究发现,在相同的Cd上,特青60天中Cd的迁移率较小,特别是在30mg/kg Cd处理下比迟心2号的低5．62％;根系细胞壁中的Cd所占的比例比迟心2号的高;特青60天根系中全硫（S）的含量比迟心2号的高,而且随着Cd处理浓度的增加,全S含量提高,但迟心2号中S含量的变化趋势与之不同;持青60天根系汁液的pH值比迟心2号的高,且汁液中Cd的含量比迟心2号的低;相关分析的结果显示;根系汁液中的Cd含量与茎叶汁液中Cd的含量呈显著的正相关关系。以上的结果可能造成了特青60天中Cd在根系内的活性和迁移能力较低,减少了Cd由根系向茎叶的迁移,从而使特青60天对Cd的抗性高于迟心2号。     

芥菜型多室油菜与甘蓝型油菜的种间远缘杂交

通过对芥菜型多室油菜与甘蓝型油菜种间杂交 以下简写为芥×甘或甘×芥 的结实性、交配性以及不同甘蓝型油菜对交配性的影响等研究发现:芥、甘正反交形成的饱满种子数较少,其形成种子的能力弱,但是芥×甘与甘×芥杂交相比,芥×甘形成饱满种子的能力较强,受精能力以及杂种胚胎的发育能力也强,在授粉后的子房发育上二者无显著差异.所以,芥菜型多室油菜与甘蓝型油菜种间杂交创建新资源时宜采用芥×甘杂交方式;不同甘蓝型油菜品种与芥菜型多室油菜正反交的结角率、受精指数、结籽指数和可交配指数均不相同,但可交配指数的变异系数最大.因此,筛选可交配性强的甘蓝型基因型应着眼于可交配指数高的甘蓝型油菜亲本材料,根据本试验结果,芥菜型多室油菜与甘蓝型油菜93-221-1杂交形成的杂种胚具有较强的可发育性.     

Differential expression of DAHP synthase and chorismate mutase isozymes during rhizogenesis of Brassica juncea cultured in vitro

The regulatory patterns of two of the enzymes of the shikimate pathway. 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHP synthase or DS. EC 4. 1. 2. 15) and chorismate motase (CM, EC 5. 4. 99. 5), were investigated using in vitro cultures of Brassica juncea at two stages, viz. undifferentiated, proliferating callus and the root-forming callus. Our studies revealed the presence of the two isozymes of DAHP synthase, DS-Mn and DS-Co. in undifferentiated callus. However, during the rhizogenesis of the callus DS-Mn was absent. Similarly, for chorismate mutase, whereas both the isozymes CM-1 and CM-2 were present in undifferentiated callus only CM-2 was detected at rhizogenesis. The possible involvement of these isozymes in callus growth and rhizogenesis is discussed.     

硅对受土壤中镉污染的白菜生长和抗胁迫能力的影响

在土培盆栽条件下,硅可提高白菜的地上部鲜重和茎鲜重,但根鲜重下降,叶鲜重略有下降。硅可抑制白菜吸收镉,在0.3、0.6、1.2mg·kg~(-1)镉水平下,施硅可显著降低白菜地上部的镉含量,并在一定程度上提高叶中过氧化物酶(POD)活性,但降低超氧化物歧化酶(SOD)活性。0.5、1.0 g·kg~(-1)硅可提高白菜的叶绿素含量和过氧化氢酶(CAT)活性,降低叶细胞膜透性,从而提高其对镉胁迫的耐受力。较高的硅浓度对植物有一定的毒性,叶绿素含量和CAT活性都下降,细胞膜透性也增加。     

Differential response of Brassica juncea cultivars to Al; consequences for chlorophyll a fluorescence,antioxidants and psb A gene

The present study was undertaken to identify the aluminum (Al) tolerant cultivar of Brassica juncea. We examined the changes in antioxidant enzymes, proline level, chlorophyll a fluorescence and psb A gene expression at vegetative and reproductive growth stages of B. juncea cultivars (Bio-902, CS-14, Pusa-Tarak and Laxmi). The selected cultivars were exposed to soil (pH 5.2) supplemented with Al (0, 50, 100 and 150?mg?kg^?1). We observed the lowest decline in photosynthetic efficiency (ΔF/Fm′), ETR, PPFD values and psb A expression to Al stress in the cultivar Bio-902 (tolerant cultivar) followed by CS-14 and Pusa-Tarak whereas the highest decline was observed in the cultivar Laxmi (sensitive cultivar). The improved performance of the cultivar Bio-902 under Al stress was accompanied by an increase in proline level, CAT and APX activities but without any increase in SOD activity. However, significant increase in SOD activity was observed in Laxmi.     

Optimization of ultrasonic-stimulated solvent extraction of sinigrin from Indian mustard seed (Brassica Juncea L.) using response surface methodology

Introduction – Sinigrin, a major glucosinolate present in Indian mustard (Brassica juncea L.) seeds as the precursor of the anticancer compound allyl isothiocyanate, shows a wide range of biological activities. It's necessary to optimize the extraction methods and conditions, in order to improve the extraction productivity and save raw material. Objective – To systemically investigate and optimize the most important factors affected the productivity of sinigrin in the process of extraction using response surface methodology. Methodology – The ranges of three main factors including the ethanol concentration, extraction time and extraction temperature were selected by the one‐factor‐at‐a‐time method. The conditions of ultrasonic‐stimulated extraction of sinigrin from defatted Indian mustard seed powder were optimized by Box‐Behnken design to obtain the maximum productivity. Result – The predicted productivity (3.81%) was obtained using 57% ethanol concentration at 81°C for 60 min, with the coefficient of the model R² > 0.96 (n = 17). The actual productivity (3.84 ± 0.02%) of sinigrin under the optimized condition was increased by 70.67% compared with the result of conventional extraction. Meanwhile, HPLC, UV and IR were applied to examine if there is a difference between the ultrasonic‐stimulated solvent extraction and conventional extraction, and the improvement of productivity of sinigrin depended on the destruction of cell wall caused by the elimination of outer pectinous material was explained by SEM and composition content analysis. Conclusion – The ultrasonic‐stimulated solvent extraction was suggested to be a promising method to improve the productivity of sinigrin. And the results demonstrated that sinigrin productivity may be related to pectinous materials existed in the seeds. Copyright © 2010 John Wiley & Sons, Ltd.     

Overexpression of Brassica juncea wild-type and mutant HMG-CoA synthase 1 in Arabidopsis up-regulates genes in sterol biosynthesis and enhances sterol production and stress tolerance

Brassica juncea 3-hydroxy-3-methylglutaryl-CoA synthase (HMGS) is encoded by four isogenes (BjHMGS1-BjHMGS4). In vitro enzyme assays had indicated that the recombinant BjHMGS1 H188N mutant lacked substrate inhibition by acetoacetyl-CoA (AcAc-CoA) and showed 8-fold decreased enzyme activity. The S359A mutant demonstrated 10-fold higher activity, while the H188N/S359A double mutant displayed a 10-fold increased enzyme activity and lacked inhibition by AcAc-CoA. Here, wild-type and mutant BjHMGS1 were overexpressed in Arabidopsis to examine their effects in planta. The expression of selected genes in isoprenoid biosynthesis, isoprenoid content, seed germination and stress tolerance was analysed in HMGS overexpressors (OEs). Those mRNAs encoding enzymes 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR), sterol methyltransferase 2 (SMT2), delta-24 sterol reductase (DWF1), C-22 sterol desaturase (CYP710A1) and brassinosteroid-6-oxidase 2 (BR6OX2) were up-regulated in HMGS-OEs. The total sterol content in leaves and seedlings of OE-wtBjHMGS1, OE-S359A and OE-H188N/S359A was significantly higher than OE-H188N. HMGS-OE seeds germinated earlier than wild-type and vector-transformed controls. HMGS-OEs further displayed reduced hydrogen peroxide (H(2) O(2) )-induced cell death and constitutive expression of salicylic acid (SA)-dependent pathogenesis-related genes (PR1, PR2 and PR5), resulting in an increased resistance to Botrytis cinerea, with OE-S359A showing the highest and OE-H188N the lowest tolerance. These results suggest that overexpression of HMGS up-regulates HMGR, SMT2, DWF1, CYP710A1 and BR6OX2, leading to enhanced sterol content and stress tolerance in Arabidopsis.     

Overexpression of<Emphasis Type="Italic">Arabidopsis</Emphasis> phytochelatin synthase (<Emphasis Type="Italic">atpcsi</Emphasis>) does not change the maximum capacity for non-protein thiol production induced by Cadmium

Phytochelatins (PCs) play an important role in heavy-metal homeostasis and detoxification. However, we previously reported that the overexpression of PC synthase inArabidopsis does not lead to increased tolerance of cadmium but, rather, plants show higher Cd sensitivity. Here, we compared the maximum capacity for non-protein thiol (NPT) production at various concentrations of Cd in order to estimate PC synthesis indirectly for both transgenic (pcs9) and wild-type plants. The pcs9 line produced the highest level of NPT when treated with 200 p.M Cd for 3 d. In comparison, the maximum productivity by the wild type was in response to 500 μM Cd. Nevertheless, the absolute amounts of NPT produced did not differ significantly between those two genotypes. Furthermore, exogenous application of 1 mM GSH did not dramatically change the capacity for either pcs9 or wild-type plants. These results suggest that Cd hypersensitivity in the transgenic pcs9 may not be caused by supraoptimal intracellular concentrations of PC, but may, instead, be due to overexpressed PC synthase itself because that enzyme can bind metals. This action, therefore, may lead to some unknown disruption in cellular metal homeostasis under Cd stress.     

Plant regeneration from peduncle segments of oil seed Brassica species: Influence of silver nitrate and silver thiosulfate

Cultured peduncle segments of B. juncea, B. campestris, B. napus, B. nigra and B. carinata produced shoot buds on Murashige and Skoog medium supplemented with benzyladenine and 1-naphthalene acetic acid. Supplementation of the media with 30 μm silver nitrate or silver thiosulfate enhanced the frequency of shoot regeneration. The regenerated shoots could be rooted at a frequency of 95% and transferred to soil where 75% survived and set seed. This revised version was published online in June 2006 with corrections to the Cover Date.     

西藏与周边地区芥菜型油菜农艺性状比较研究

运用典范相关分析方法,对西藏及周边地区芥菜型油菜的产量性状、主茎性状、分枝性状、角果性状等4组性状间的典范相关关系进行了比较研究.结果表明:(1)所研究的18个性状中,西藏芥菜型油菜的平均数为周边地区芥菜型油菜的1.88倍,总体变异系数比周边省份芥菜型油菜高10.22％;(2)西藏芥菜型油菜单株产量主要取决于每株角果数和千粒重,而周边省份芥菜型油菜单株产量主要取决于每株有效角果数,中国周边国家芥菜型油菜单株产量主要取决于每株有效角果数和千粒重;(3)影响西藏芥菜型油菜产量性状最重要的因素是角果性状,其次是分枝性状和主茎性状,而影响周边省份和中国周边国家芥菜型油菜产量性状最重要的因素则是主茎性状,其次是分枝性状和角果性状.     

Molecular markers linked to white rust resistance in mustard Brassica juncea

 White rust, caused by Albugo candida (Pers.) Kuntze, is an economically important disease of Brassica juncea (L.) Czern. and Coss mustard, particularly in India. The most efficient and cost-effective way of protecting mustard plants from white rust disease is through genetic resistance. The objective of this study was to identify RAPD markers for white rust resistance in an F₁-derived doubled-haploid (DH) population originating from a cross between white rust-susceptible and white rust-resistant breeding lines of B. juncea from the canola-quality B. juncea breeding project of the Agriculture and Agri-Food Canada-Saskatoon Research Centre. The DH population was used to screen for RAPD markers associated with white rust resistance/susceptibility using bulked segregant analysis. Two markers, WR2 and WR3, linked to white rust resistance, flanked the resistance locus Ac2 ₁ and were highly effective in identifying the presence or absence of the resistance gene in the DH population. These two markers were shown to be specific to the Russian source of white rust resistance utilized in this project. It is concluded that the availability of these RAPD markers will enhance the breeding for white rust resistance in B. juncea. Received: 17 December 1997 / Accepted: 7 April 1998     

Callus Formation and Morphogenesis of Cultured Mustard (Brassica juncea) Protoplasts

Protoplasts from mustard seedling cotyledons were suspended in Nitsch medium and cultured at 26℃ under low intensity illumination. When colonies were observed in the cultures, fresh medium (3% sucrose replaced mannitol) was added by equalvolume. When protoplasts developed into small callus the cultures were transfered to modified MS agar medium, in which some of them produced roots while an others gave rise to green spots. Shoot-differentiation experiment is in progress.