Effect of Combined Use of Boron and Manganese Fertilizers on the Nutritional Quality and Physiological Indices of Brassica campestris L.ssp.chinensis L.var.utilis

[Objective] The purpose of this study is to determine the effects of com-bined use of boron and manganese fertilizers on the nutritional quality and physio-logical indices of Brassica campestris. [Method] In the nutrient solutions for growing B. campestris by hydroponics, boric acid and manganese sulfate were added at 0.5, 2.5, and 7.5 mg/L respectively. Another treatment without boron and manganese was prepared as the control. Quality and physiological indices of B. campestris in the 10 treatments were measured. [Result] Boron and manganese shared obvious in-teraction in improving the quality and physiological indices of B. campestris. To cul-tivate B. campestris with high quality and strong resistance, the optimum concentra-tions of boron and manganese in the nutrient solution should be 2.5 mg/L boric acid and 2.5-7.5 mg/L manganese sulfate. [Conclusion] The findings wil provide refer-ence for studying effects of trace elements on nutrient composition of vegetables.

Effect of Nitrogen and Sulfur Supply on Glucosinolates in Brassica campestris ssp.chinensis

Glucosinolates （GSs） are a group of plant secondary metabolites containing abundant nitrogen （N） and sulfur （S） mainly in Brassica and have the beneficial effects on human health including anti-carcinogenic, cholesterol-reducing and other pharmacological effects. The objective of this study was to investigate the effect of different concentrations of N （5, 10, and 20 mmol L-a, denoted by N5, N10 and N20） and S （0,5, 1, and 2 mmol L^-1, denoted by S0.5, S1 and S2） on the yield and GSs in pakchoi （Brassica campestris L. ssp. chinensis var. communis） in hydroponics. Results showed that N10 and N20 significantly enhanced the yield compared with N5, however, N20 had a negative effect relative to N10. Only with N10 and N20 low S supply （S0.5） reduced the yield. The concentrations of aliphatic GSs, aromatic GS and total GSs were enhanced by N5 and indolyl GSs were enhanced by N20. S2 enhanced the concentration of individual GS and total GSs. The concentrations of indolyl GSs were maximized in N20S2 treatment, whereas the highest concentrations of aliphatic GSs, aromatic GS and total GSs were found in N5S2 treatment. Effects of N and S on aliphatic GSs were higher than on indolyl GSs. The results suggest that the accumulation of aliphatic GSs and aromatic GS could be enhanced by low N and high S and restricted by high N while that of indolyl GSs could be enhanced by high N and high S.

Nitrogen Dioxide-Induced Responses in Brassica campestris Seedlings: The Role of Hydrogen Peroxide in the Modulation of Antioxidative Level and Induced Resistance

This article investigates the responses of Brassica campestris seedlings to an acute level of nitrogen dioxide （NO2） exposure in a plant growth chamber, and examines whether pretreating plants with hydrogen peroxide （H2O2） will alleviate NO2-caused injury. Twenty-eight-day-old B. campestris plants sprayed with 10 mmol L^-1 H2O2 aqueous solution （corresponding to approximate 1.0 mg H2O2 per single plant） were exposed to different concentrations of NO2 （0.25, 0.5, 1.0, and 2.0 μL L^-1, respectively） for 24 h under controlled environment. To measure the plant biomass, the plants were fumigated with the same NO2 concentrations as mentioned above for 7 h per day （8.00-15.00） for 7 days. As a control, charcoal filtered air alone was applied. Data were collected on plant biomass, total chlorophyll, photosynthetic rate, stomatal conductance, nitrate and nitrate reductase （NR）, antioxidative enzymes, ascorbate （ASA）, and malondialdehyde （MDA）, immediately after exposure. The results showed that exposure to a moderate dose of NO2 （e.g., 0.25 μL L^-1） had a favorable effect on plants, and the dry weight of the above-ground part increased, whereas the exposure to high NO2 concentrations （e.g., 0.5 μL L^-1 or higher） caused a reduction in the plant biomass and the total chlorophyll, when compared with the control. In addition, at 0.5 μL L^-1 or higher NO2 concentrations, prominent increases in the MDA level and superoxide dismutase （SOD） and NR activities were observed. Exposure to 1 μL L^-1 and higher NO2 resulted in necroses appearing on older leaves, and an increase in catalase （CAT） activity, decrease in ASA content, increased accumulation of NO3^-, and reduction in photosynthesis, when compared with the controls. No changes were detected in stomatal conductance under NO2 fumigation. The pretreatment with 10 mmol L^-1 H2O2 alleviated significantly NO2- caused biomass decrease and photosynthetic inhibition when compared with H2O2-untreated plants. Under NO2 fumigation, further induction in SOD and CAT activities occurred in H2O2 treated plants when compared with H2O2- untreated plants. The effect of NO2 on the ASA and MDA contents was also absent in H2O2-treated plants. However, the H2O2 treatment did not alter the nitrate content and NR activity in plants under NO2 fumigation. The H2O2 treatment caused a lower rate of stomatal conductance. Taken together, these data suggest that fumigation with an acute level of NO2 causes oxidative damage to B. campestris seedlings. The H2O2 pretreatment markedly protects plants against NO2 stress and this may be associated with inducible antioxidative level. NO2 fumigation contributes, at least in part, to the enhanced levels of nitrate in B. campestris leaves.

Preliminary Study on Tissue Culture Technique of Brassica campestris L. ssp. chinensis (L.) Makino var. utilis

In order to establish a rapid propagation system for Brassica campestris L. ssp. chinensis var. utilis, the anther and cotyledon-cotyledonary petioles were used as explants to conduct tissue culture research. The results showed that not bloomed flower buds with higher styles than petals were appropriate for anther cul- ture. Moreover, most microspores were at the mid-late uninucleate stage at that time. The pollen germination rates of Brassica campestris L. ssp. chinensis var. u- tills were low and the germination rates in autumn and winter were higher than that in summer. The induction medium of anther callus of Brassica campestris L. ssp. chinensis var. utilis was MS＋1.0 mg/L KT＋1.0 mg/L 2, 4-D＋3% sugar＋6 g/L agar＋ 8% coconut milk and the induction media of adventitious bud were MS＋2.0 mg/L 6-BA＋ 0.5 mg/L NAA＋I.0 g/L activated carbon＋2% sugar＋6 g/L agar or MS＋2.0 mg/L ZT＋ 0.5 mg/L IAA＋0.5 g/L AgNO3＋1.0 g/L activated carbon＋2% sugar＋6 g/L agar. The induction percentage of adventitious bud by anther culture was 36.7%. Browning phenomenon appeared during the culture of adventitious bud and regeneration plant could not be formed. The plant regeneration rate reached 80% when cotyledon- cotyledonary petioles were used as explant.

Differential expression of salt tolerance related genes in Brassica campestris L. ssp. chinensis (L.) Makino var. communis Tsen et Lee

We examined salt tolerance responsive genes in Pak-choi under salt stress and analyze their potential function. The LRNA differential display was used to screen the transcript derived fragments （TDFs） related to salinity tolerance in tolerant and Loderately tolerant Pak-choi germplasm. Seventy-eight primer combinations generated 101 differential eDNA fragments, which ere divided into 10 expression types. Seven cDNA sequences （GenBank accession Nos. DQ006915-DQ006921） obtained and ,~quenced were highly homologous to some known expression genes or the genes related to the signaling pathways in plants under ifferent abiotic stress.

正交设计优化菜薹ISSR反应体系研究

菜薹原产中国,是华南地区重要的特产蔬菜之一。目前,分子标记技术已广泛应用于多种作物研究,但在菜薹上的应用研究刚刚起步。ISSR主要是由单一引物且以重复序列为主要引物序列的PCR标记,其反应条件受模板DNA、Taq DNA聚合酶、Mg2+、dNTP和引物等因素的影响。正交试验设计能明确各因素间的互作效应,建立最优化的反应体系。本研究利用正交试验设计,从Taq DNA聚合酶、dNTP、引物、Mg2+4种因素3个水平对菜薹ISSR反应体系进行了优化,确立了适合菜薹的ISSR反应体系并在7个菜薹品种中进行了验证。在20μl反应体系中,含Taq DNA聚合酶1U、dNTP250μmol/L、引物0.25μmol/L、1×PCRbuffer、Mg2+2.5mmol/L、模板DNA30ng。通过梯度PCR测验,确定了适宜的退火温度。这一体系的建立为今后利用ISSR技术进行菜薹种质资源分类、遗传图谱构建和基因定位奠定了技术基础。

不同土壤类型对红菜薹品质的影响

为摸清历史上与洪山区同属武昌县的江夏区种植洪山菜薹的可行性,在位于江夏区法泗镇的公司甲、位于江夏区郑店街的公司乙、洪山菜薹原产地进行了不同土壤类型种植洪山菜薹品质对比试验。对6个不同土壤样品进行检测,在红菜薹(Brassica campestris L.ssp Chinensis L.var.utilis Tsen et Lee)种质和栽培管理相同条件下,分别测定对应土壤的红菜薹主薹和侧薹品质。结果表明,原产地种植的洪山菜薹主薹品质最好,公司甲黄土种植的洪山菜薹侧薹品质最好;在品种、田间管理相同的条件下,土质是关键所在;在江夏区搞好洪山菜薹示范生产,延伸产业链是可行的。

一种红菜薹新病害的病原鉴定及生物学特性研究

2010-2011年在湖北省红菜薹(Brassica campestris L.ssp.chinensis L.var.utilis Tsen et Lee)栽培中发现一种新病害,并从该红菜薹新病害发病叶片分离得到了病原菌hctyk1,观察该病原菌的孢子形态,分析了病原菌的ITS序列,研究了病原菌的生物学特性。结果表明,该病原菌为链格孢[Alternaria alternata(Fr.:Fr.)Keissler],病原菌r DNA-ITS区序列已在Gen Bank上登录(登录号:JQ885954)。该菌菌丝生长致死温度为50℃;适宜pH为8;在供试的几种碳源、氮源中,最适的碳源是葡萄糖,最适的氮源是酵母浸出液。

红菜薹废弃物还田对四季豆产量和养分累积的影响

通过田间试验研究了红菜薹(Brassica campestris L. ssp. chinensis var. utili Tsen et Lee)废弃物还田以及化肥减施对四季豆产量和养分累积的影响。结果表明,在常规施肥的基础上,红菜薹废弃物半量和全量还田四季豆产量分别增加了13.41%和29.43%,与红菜薹废弃物全部还田相比,全部还田+化肥用量减施30%使得四季豆产量降低了25.41%;与单施化肥相比,全部还田+化肥用量减施30%处理四季豆生物量增加了9.38%。红菜薹还田氮素、磷素和钾素累积量分别较单施化肥处理增加了18.72%~32.55%、6.35%~38.22%和5.73%~41.87%,且富集的氮、钾主要存在茎、叶片和果实中。

苋菜-小白菜混作对小白菜镉与硝酸盐复合污染的影响

采用酸性黄棕壤和石灰性潮土进行连续两季盆栽试验,研究了镉与硝酸盐复合污染条件下苋菜(Amaranthus mangostanus L.)与小白菜[Brassica campestris L.ssp.chinensis(L.)Makino.var.communis Tsen et Lee]混作对小白菜品质及产量的影响。结果表明,在Cd 2.0 mg/kg与N 0.2 g/kg的交互作用下,苋菜-小白菜混作显著降低(P<0.05)了小白菜镉与硝酸盐的含量,黄棕壤第一季、第二季及潮土第一季、第二季收获的小白菜硝酸盐含量分别降低了19.87%、17.31%、27.00%、32.45%;小白菜的镉含量分别降低了23.33%、22.58%、5.82%、7.69%。同时,苋菜-小白菜混作也能提高小白菜的品质及产量,黄棕壤第一季、第二季及潮土第一季、第二季与对照相比,小白菜的维生素C含量分别提高了47.92%、22.66%、29.66%、92.26%,差异均达显著水平(P<0.05);可溶性蛋白质含量分别提高了27.44%、45.29%、39.50%、50.00%,差异均达显著水平(P<0.05);叶绿素含量分别提高了11.68%、19.38%、1.18%、7.47%;可溶性糖含量分别提高了1.34%、31.05%、321.05%、154.55%;小白菜的产量分别提高了2.26%、3.48%、1.89%、19.53%。对小白菜品质及产量而言,均有不同程度的提高,尽管有的差异不显著,但仍有提高的趋势,这对于试验范围内的植物修复而言,均有积极意义。

NaCl胁迫对不同生育期红菜薹的影响

研究了NaCl胁迫对不同生育时期红菜薹[Brassica campestris L.ssp.chinensis(L.)Makino var.utilis Tsen et Lee]的影响。结果表明,在种子萌发期,300 mmol/L的NaCl处理显著降低了红菜薹种子的发芽率、发芽势和发芽指数;在幼苗期,不同浓度的NaCl处理对红菜薹的地上部鲜重、根鲜重、根干重、丙二醛含量、根系活力均无显著影响,在300 mmol/L的NaCl胁迫下红菜薹的地上部干重反而显著增加;在成株期,不同浓度的NaCl处理对红菜薹的叶片数、主薹高度、主薹茎粗、侧薹数、单株产量、丙二醛含量、根系活力均无显著影响。

轮作和有机肥对连作小白菜生长及土壤微生物特性的影响

为研究轮作和有机肥处理对连作土壤的修复作用,以小白菜(Brassica campestris L.ssp.chinensis Makino var.communis Tsen et Lee)连作土壤为对照,比较玉米轮作,玉米秸秆还田,玉米秸秆有机肥等对连作土壤酶活性、微生物活性、活性有机碳含量及小白菜光合速率、根系活力、根系生长的影响。结果表明,轮作和有机肥处理不同程度地增加了连作小白菜光合速率、干物重、根系活力和根系ATPase活性,降低了根系MDA含量,同时提高了连作土壤酸性磷酸酶、脲酶、转化酶活性和微生物对碳源的利用能力,其中长期轮作及施用发酵有机肥的促进作用较大。相关分析表明,土壤酶活性、根系活力、活性有机碳含量、碳源利用AWCD值之间存在极显著正相关关系,说明采用合适的有机肥投入方式可以缓解连作对作物生长造成的抑制。

不同青菜品种吸锌能力差异及与根系分泌物的关系

青菜 [Brassicacampestrisvar chinensis(L )Makino]不同品种对土壤中锌的累积吸收有明显差异[1] ,我们推断根系分泌物在青菜吸收锌的过程中可能起着重要作用。因此在本项研究中 ,对 2个不同锌利用效率的青菜品种的根系分泌物组成、数量及其与锌吸收的关系进行了探讨。结果表明 ,2种锌水平下 ,五月慢根系分泌物中低分子量有机酸及氨基酸的总量均明显高于黑油白菜 ,缺锌时品种间差异达 0 0 5显著水平 ;缺锌条件下 ,五月慢根系分泌物中草酸、丙氨酸的含量同样显著高于黑油白菜。五月慢在缺锌时有较高的根系分泌物总量及其草酸、丙氨酸含量都显著增加 ,这可能是五月慢具有较高累积吸锌量及锌利用效率的重要原因之一。

乌塌菜ISSR-PCR反应体系的建立及优化

为获得乌塌菜ISSR-PCR的最佳反应体系,采用单因素浓度梯度试验和正交优化设计相结合的方法,研究了引物浓度、dNTP浓度、Mg2+浓度、Taq DNA聚合酶用量对PCR反应的影响。并在此基础上对退火温度和循环数进一步优化,最终确立了适合乌塌菜ISSR-PCR反应的最佳体系和程序。即20μL PCR反应体系含:30 ng模板DNA,0.50μmol.L-1引物,0.25 mmol.L-1 dNTP,1 mmol.L-1 Mg2+,1.0 U Taq DNA聚合酶。PCR扩增程序为:94℃预变性3 min;94℃变性30 s,50℃退火1 min,72℃延伸90 s,35个循环;72℃延伸7 min,4℃保存。这一优化的ISSR-PCR反应体系的建立为今后利用ISSR技术对乌塌菜进行种质资源的分类鉴定奠定了基础。

菜心基因组DNA提取及RAPD反应体系的优化

用改良的CTAB法提取菜心基因组DNA,并利用紫外分光光度法测定其纯度和含量,用琼脂糖凝胶电泳法检测DNA的降解状况和纯度．结果表明:此法提取的DNA具有典型的天然DNA分子的标准紫外吸收光谱特点, 且适于进行RAPD分析．RAPD分析的优化反应体系为:25μL的反应液中含有0．8 U的Taq酶,0．28 μmol／L primer,30 ng的DNA,2．0 mmol／L Mg2+,0．20 mmol／L dNTPs．PCR扩增循环为94℃、5 min;94℃、1 min; 36℃、1 min;72℃、2 min,37个循环;72℃延伸10 min．

27个菜心品种遗传多样性的ISSR分析

利用ISSR技术对27个菜心品种的遗传多样性进行了分析.结果表明,12条引物共检测到103个可重复位点,其中多态位点58个,多态位点百分率(P)为56.31%.平均多样性指数(I)为0.229,遗传相似系数(H)为0.144,观察等位基因数(Na)和有效等位基因数(Ne)分别为1.563和1.225,遗传距离(GD)的范围在0.029～0.344之间,表明菜心遗传多样性较低.经UPGMA聚类分析,将27个菜心品种分为六大类群.

TA29-barnase基因转化菜心

利用根癌农杆菌导入法,以菜心带柄子叶为外植体,对TA29-barnase基因转化菜心进行研究。获得转化植株,进行PCR、Southern blotting杂交和半定量RT-PCR检测,表明目的基因已经整合到转化植株中,并且目的基因在转基因植株花蕾中得到表达,但是表达水平在不同转基因植株间存在差别;转基因植株开花后,均表现雄性不育,不能产生花粉或产生没有活力的少量花粉,自交不能结实;用未转化植株正常花粉对雄性不育植株进行授粉,能够正常结实;保持系(未转化植株)与不育株杂交后代中雄性不育株与可育株的比例为1:1,在杂交后代植株子叶期,喷洒10mg/L的PPT可以完全杀死可育株;利用其他菜心品种为父本与不育株进行杂交,获得的F1植株在生长势和产量方面表现优势,表明开展菜心优势育种具有一定的潜力。

我国乌塌菜研究进展

在植物学分类上,乌塌菜〔Brassica campestris L.ssp.chinensis(L.)Makino var.rosularis Tsenet Lee〕属于十字花科芸薹属白菜亚种的一个变种,别名塌菜、塌棵菜、塌地菘、太古菜、黑菜等,其营养价值较高,含有丰富的矿物质和维生素。本文综述了我国乌塌菜种质资源收集鉴定及主要种类、新品种选育、育种技术、栽培技术、生理相关研究以及分子技术等方面取得的进展和研究成果,分析了目前我国乌塌菜研究中存在的主要问题,同时对未来工作进行了展望。

菜心ISSR-PCR反应体系的优化

以菜心（Brassica campestris L．ssp．Chinensis Var．utilis Tssen．et Lee．）为材料，对影响ISSR-PCR扩增结果的因素如Mg^2＋、Taq DNA聚合酶、dNTPs、Primer、模版DNA的浓度及引物退火温度、延伸时间和循环次数进行了探讨，确立了适合菜心ISSR-PCR分析的最佳反应体系和PCR扩增参数：在25pL反应体系中含10×buffer 2．5μL，2．0mmol／LMg^2＋，0．5U Taq DNA聚合酶，0．2mmol／LdNTPs，0．5μmol／L引物，30ng模板DNA．PCR扩增程序：94℃预变性3min；94℃变性1min，49．7～56℃退火（退火温度随引物不同而定）1min，72℃延伸45s，40个循环；72℃延伸5min．

菜薹细胞质不育系小孢子发生的细胞形态学研究

菜薹雄性不育系小孢子发生的细胞形态学观察表明，孢原细胞分化期之前败育的花占５９．６％，不形成花粉囊；有花粉囊发育的花占４０．４％，其中绝大多数花中只有一个雄蕊能形成一个体积很小的花粉囊，雄性不育系小孢子发育在小孢子母细胞期、四分体及单核小孢子期均有败育，败育方式有绒毡层细胞异常肥大，挤压小孢子母细胞；减数分裂异常，无胞质分裂而形成巨细胞；单核小孢子粘连成花粉块，异常膨大等。还有雄蕊雌蕊化等花器异常现象。