Transmission of the agent causing a melon yellowing disease by the greenhouse whitefly Trialeurodes vaporariorum in southeast Spain

The agent causing a yellowing disease of melon (Cucumis melo), which results in severe losses in crops under plastic on the coastal plains of southeast Spain, was shown to be transmitted in a semipersistent manner by the greenhouse whitefly (Trialeurodes vaporariorum Westwood). The agent was transmitted by grafting, but not by mechanical inoculation or through seeds. The agent was acquired in the minimum period tested (2 h) and could infect plants in an infection feeding interval of 6 h. Capsella bursa-pastoris, Cucumis melo, C. sativus, Cucurbita moschata, Cichorium endivia, Lactuca sativa andTaraxacum officinale were found susceptible.Results suggest that the yellowing disease affecting melon crops in the southeast of Spain is due to a pathogen similar to beet pseudo yellows virus, but this has to be confirmed by serology.     

黄瓜全雌性基因连锁的AFLP和SCAR分子标记

 本研究以全雌品种‘戴多星’自交系和弱雌品种‘北京截头’自交系为双亲杂交获得F₁ ,然后得到F₂ 性型分离群体, 利用分离群体分组分析法(Bulked Segregant Analysis, BSA) 构建全雌和弱雌两个基因池, 筛选了64对AFLP选择性引物EcoR I-NN +Mse I-NNN组合, 发现EcoR I-TG +Mse I-CAC引物组合在全雌基因池中扩增出一条分子量为234 bp的特异带。经F₂ 代单株验证, 该特异条带能在全雌单株中稳定出现。以MAP MAKER (Version 310) 软件分析, 该标记与全雌性位点的连锁距离在617 cM。命名该连锁标记为TG/CAC₂₃₄。将该特异条带回收、克隆、测序, 设计特异SCAR引物, 再对F₂ 代单株基因组DNA进行扩增, 仅在全雌单株中扩增出1条分子量为166 bp 的特异带, 表明已成功地将与黄瓜全雌性连锁的AFLP标记转化为操作简便、表现稳定的SCAR标记, 该标记命名为SA₁₆₆。     

从甜瓜子叶中提取总DNA

用分子标记方法检测甜瓜杂交种子纯度,如果从真叶中提取DNA,检测周期至少需要15d。为了缩短检测周期,我们分别以甜瓜的成熟干种子、吸胀水后“露白”种子、3d龄刚转绿子叶、6d龄子叶、9d龄子叶和12d龄子叶为材料,进行了提取总DNA的研究。结果表明:除了干种子和吸胀水后“露白”种子外,其他的材料都可以提取到DNA,但是DNA的质量因子叶日龄不同而存在很大的差异。不同日龄的子叶中,以3d龄子叶提取的DNA质量好。子叶6d龄时,提取的DNA质量次于3d龄,少量DNA开始出现降解,以后随着子叶日龄的增加,DNA降解加重。另外,与以真叶为材料提取的DNA样品相比较,子叶DNA样品中的蛋白质等杂质含量高,应增加氯仿抽提纯化次数。     

Further suppression of<Emphasis Type="Italic">Botrytis cinerea</Emphasis> disease in cucumber seedlings by chitosan-copper complex as compared with chitosan alone

Chitosan-copper complex compared with chitosan alone enhanced suppression ofBotrytis cinerea rot development on four-true-leaf cucumber seedlings in controlled growth chambers. This paper constitutes the first report of such enhancement. The optimal concentrations for the most effective suppression ofBotrytis development were 0.2 gl ⁻¹ chitosan and 1.6 mmole copper. After 12 days’ incubation, marked and significantly better disease suppression was obtained with chitosan-copper complex (75% suppression) than with chitosan alone. The chitosan-copper complex could be a very promising decay control agent for use in both conventional and organic agriculture. http://www.phytoparasitica.org posting Dec. 18, 2002.     

黄瓜单体异附加系的筛选与观察

 将甜瓜属异源三倍体与栽培黄瓜‘北京截头’杂交，经胚胎拯救获得了两个2n=15 (14C+1H)的单体异附加系，编号分别为02—17和02—39。02一l7植株叶形为戟形，02—39为深陷的掌状形，并且果刺均为白色，不同于异源三倍体的掌状心脏形和黑刺，果形比异源三倍体黄瓜长而更接近普通黄瓜。细胞学研究发现，单体异附加系在终变期和中期I染色体构型主要是由7个二价体和1个单价体构成，出现三价体的频率为4％ ～5％ ，中期Ⅱ不均等分裂形成不同的两极，一极染色体为8条，另一极为7条。RAPD分析证明它们分刖附加了野生种Cucum／s hystrix的染色体。     

Transgenic cucumber expressing an endogenous class III chitinase gene has reduced symptoms from Botrytis cinerea

A class III chitinase gene (CHI2) is induced in cucumber plants (Cucumis sativa L.) in response to infection by pathogenic microorganisms. Infection of Botrytis cinerea, causal agent of gray mold disease on cucumber, also induces CHI2 expression. To investigate whether CHI2 is involved in resistance to gray mold disease, transgenic cucumber plants were produced to overexpress the CHI2 gene. One line was analyzed in detail in terms of disease resistance. The transgenic cucumber plant (CC2) constitutively expressed CHI2 and reduced the symptoms of B. cinerea for 4 days after inoculation compared with nontransgenic plants. However, this inhibitory effect was not absolute, and CC2 eventually developed serious disease symptoms. Chitinase activity of the crude extract from CC2 leaves was higher than that from nontransgenic plants. A high-molecular-weight fraction containing CHI2 from CC2 leaves had fungistatic activity against B. cinerea. Interestingly, the low-molecular-weight fraction from CC2 leaves with CHI2 removed also had fungistatic activity against B. cinerea. Not only the introduced chitinase activity but also the endogenous defense reactions activated by overexpression of CHI2 may be involved in the enhanced gray mold disease resistance in CC2.     

转基因甜瓜植株的获得及其抗病性

甜瓜（Ｃｕｃｕｍｉｓ ｍｅｌｏ）子叶外植体能被含双元载体的根瘤农杆菌菌不妨功体内含１个ＮＰＴ－Ⅱ基因（新霉素磷酸基转移酶Ⅱ基因）,以供选择卡妹妹纱抗性的转化甜 １个ＷＭＶ－２ ＣＰ基因（西瓜花叶病毒２号外壳蛋白基因）。卡那霉素抗性植株经ＰＣＲ－Ｓｏｕｔｈｅｒｎ杂交试验表明,外源的ＷＭＶ－２ ＣＰ基因确已通过农杆菌介导的转化途径导入甜瓜细胞。目前转基因甜瓜已开花结果。攻毒试验表明,与对照相比,转基因     

黄瓜种质资源遗传亲缘关系的RAPD分析

利用ＲＡＰＤ技术对多个生态型的黄瓜材料的遗传亲缘关系进行了研究。从２００个１０碱基随机引物中筛选出２０个用于ＰＣＲ反应,３９．２％的扩增条带表现多态性;每个生态型品种都具有其特有的扩增（缺失）带以区别其他生态型品种;聚类分析将供试材料分为３大类群：华北类群、华南类群和欧洲温室类群。从分子水平验证了传统的黄瓜地域分类标准及黄瓜是遗传基础狭窄的蔬菜作物。对利用ＲＡＰＤ技术进行亲缘控制研究的可能性进行了探讨。     

甜瓜多聚半乳糖醛酸酶反义基因植物表达载体的构建及其转化烟草的研究

用限制性内切酶从目的基因供体质粒pBI-aPG上切下大小约2.3kb的目的基因,将它定向连接在受体质粒pCAMBIA2301载体上,构建成含有GUS基因和NPTⅡ基因的甜瓜多聚半乳糖醛酸酶反义基因植物表达载体pCB-aPG。采用直接转化法将pCB-aPG导入根癌农杆菌菌株LBA4404,采用该菌株对普通烟草进行了遗传转化研究。在Kanamycin选择压力下获得的烟草转化不定芽和完整植株,经过GUS基因组织化学法检测以及PCR方法鉴定,证实了该反义基因已导入烟草基因组中。此项研究为下一阶段用该反义基因转化甜瓜品种以改良甜瓜果实耐贮运性打下基础。