Multi#ex—CAPS技术及其在番茄遗传鉴定中的应用

 ：应用Multiplex—PCR和CAPS(Cleaved amplified polymorphic sequence)原理研究建立了Mutiplex—CAPS技术。该技术包括：(1)在PCR扩增反应体系加入2对、3对或多对核苷酸引物，经30～40个扩增循环后得到每对引物的特异扩增片段；(2)应用同一种限制性内切酶消化这些不同引物经扩增后获得的特异片段；(3)琼脂糖凝胶电泳溴化乙锭染色检测酶切片段长度的多态性。利用该技术对番茄(Lycopersicon es—cdentum)TA517及其近等基因系的分析表明，Multiplex—CAPS能同时检测2个、3个或多个CAPS标记的多态性，其效果等同于每个CAPS标记的多态性的叠加，重复性好，分析效率高，降低成本数倍。     

利用PCR技术同时鉴定番茄抗根结线虫和抗斑萎病毒基因

 利用同一PCR反应体系，对分别与番茄抗根结线虫的 基因和抗斑萎病毒(1w V)的Sw一5基因紧密连锁的SCAR标记进行了同时扩增筛选，扩增的特异性片段与单引物扩增片段完全吻合，其中与基因紧密连锁的SCAR1标记为共显性标记，抗感试材均产生750 bp的特异片段，纯合和杂合抗病基因型试材存在 I酶切位点，酶切后分别产生了570 bp、160 bp和750 bp、570 bp、160 bp的不同特异性片段，而感病基因型试材无 I酶切位点；与Sw一5基因紧密连锁的SCAR2标记为显性标记，只有抗病试材扩增出400 bD的特异性片段。经反复验证，结果稳定准确可靠，可用于在同一PCR反应体系中对两个抗病基因进行同时筛选鉴定。     

Biological and molecular characterization of Tomato spotted wilt Virus in Israel

Received April 24, 1997; received in final form June 29, 1997. Symptoms resembling tomato spotted wilt virus (TSWV) infections were documented among ornamental and vegetable crops in commercial greenhouses and open fields in Israel. Plants exhibiting these symptoms were collected from January 1992 to December 1996. Among cultivated plants analyzed for TSWV by enzyme-linked immunosorbent assay (ELISA), 19 species representing five families were found to be infected; natural infection was also recorded in six plant species of weeds. Virus identity was characterized by host range, serology and electron microscopy. Serological reaction with the isolates, found in Israel, using antisera from different sources as well as the sequence analysis of the nucleocapsid gene, demonstrated that the Israeli isolates of TSWV are a member of tospovirus serogroup I, type I (BR-01 strain). No virus transmission was found in seeds collected from virus-infected vegetable and ornamental crops. A non-radioactive molecular probe derived from the cloned nucleocapsid isolate enables specific detection of the virus in crude sap from infected plants. The detection of TSWV in Israel constitutes a severe potential threat to the ornamental and vegetable industry.     

三种外检植物病毒的灭活处理

番茄环斑病毒（ＴｍＲＳＶ），烟草环斑病毒（ＴＲＳＶ），南芥菜花叶病毒（ＡｒＭＶ）的病汁液和ＰＥＧ粗提纯液，经适宜温度或甲醛处理，均丧失侵染性而有抗原性。ＴｍＲＳＶＰＥＧ粗提纯液经６０℃（水浴）处理１０分钟或２８℃７天，ＴＲＳＶ粗提纯液置２５℃一个月，Ａｒ－ＭＶ在４０℃７天条件下均可做为阳性对照的灭活处理的最佳方案。可保存抗原性在７～１２月以上。为了防止危险性检疫病毒的侵入，对入境种苗进行检疫，需建立快速、准确、标准化检疫程序，而在血清学快速诊断试验中，提供阳性对照，对提高判断准确率是必要的。因此，为了解决在诊断试剂中提供阳性对照但又不能使其检疫性病毒人为扩散这一重要问题，我们在研究三种外检病毒（ＴｍＲＳＶ，ＴＲＳＶ，ＡｒＭＶ）的检验技术的同时，又进行了一些灭活试验，用化学和物理方法钝化病毒，使其失去侵染性而保持抗原性，并应用于酶联诊断试剂盒中，本文报道了初步试验结果     

中国18 省市番茄晚疫病菌生理小种的鉴定

 利用5 个含有不同单显性抗番茄晚疫病基因的番茄材料Ts19、Ts33、W1Va700、LA1033 和L3708为鉴别寄主, 对我国18 省、市、自治区的番茄晚疫病菌201 个纯化分离物进行了生理小种鉴定。结果共鉴定出8 个小种, 即生理小种T0、T1、T1 ,2 、T1 ,2 ,3 、T1 ,2 ,3 ,4 、T1 ,4 、T1 ,2 ,4 和T3。其中, 小种T1 和T1 , 2是主流小种, 地理分布最广和发生频率最高, 分别在13 和11 个省市出现, 占样本总数的28.8 %和28.4 %;其次是小种T0 和T1 ,2 ,3 , 分别在8 和7 个省市出现, 分别占1819 %和819 %; 再其次是小1 ,2 ,3 ,4 , 在4个省市有发生, 占710 %; 小种T1 ,2 ,4 、T1 ,4 和T3 分布均少, 仅在1～2 个省市发生, 分别占3.0 %、2.5 %和2.5 %。这是我国的首次报道, 为番茄晚疫病的抗源材料筛选和抗病育种提供了病理学依据。     

京郊保护地秋季番茄的氮素供应及利用

研究了氮素供应水平对保护地番茄产量、养分吸收和氮素利用的影响。结果表明,氮素供应量(N)为225kg·hm~(-2)时番茄的产量最高,但随着施氮量的增加,番茄产量反而有所下降。增施氮肥对番茄地上部N、P、K的吸收没有明显的影响。在京郊传统生产条件下,保护地番茄生产相对合适的氮素供应水平(播前根层起始土壤无机氮含量+化学氮素用量)为510 kg·hm~(-2),但如果考虑番茄生长期间土壤氮素的矿化作用,番茄生产的氮素供应水平应为584kg·hm~(-2)。     

Trichoderma Biocontrol of Colletotrichum acutatum and Botrytis cinerea and Survival in Strawberry

Trichoderma isolates are known for their ability to control plant pathogens. It has been shown that various isolates of Trichoderma, including T. harzianum isolate T-39 from the commercial biological control product TRICHODEX, were effective in controlling anthracnose (Colletotrichum acutatum) and grey mould (Botrytis cinerea) in strawberry, under controlled and greenhouse conditions. Three selected Trichoderma strains, namely T-39, T-161 and T-166, were evaluated in large-scale experiments using different timing application and dosage rates for reduction of strawberry anthracnose and grey mould. All possible combinations of single, double or triple mixtures of Trichoderma strains, applied at 0.4% and 0.8% concentrations, and at 7 or 10 day intervals, resulted in reduction of anthracnose severity; the higher concentration (0.8%) was superior in control whether used with single isolates or as a result of combined application of two isolates, each at 0.4%. Only a few treatments resulted in significant control of grey mould. Isolates T-39 applied at 0.4% at 2 day intervals, T-166 at 0.4%, or T-161 combined with T-39 at 0.4% were as effective as the chemical fungicide fenhexamide. The survival dynamics of populations of the Trichoderma isolates (T-39, T-105, T-161 and T-166) applied separately was determined by dilution plating and isolates in the mixtures calculated according to the polymerase chain reaction (PCR) using repeat motif primers. The biocontrol isolates were identified to the respective species T. harzianum (T-39), T. hamatum (T-105), T. atroviride (T-161) and T. longibrachiatum (T-166), according to internal transcribed spacer sequence analysis.     

早熟番茄新品种津粉207的选育

津粉207是利用自交系09-163为母本、10-202为父本杂交育成的番茄杂交一代品种。无限生长类型,耐低温,早熟、果实粉红、果形美观,单果质量在220 g以上,产量高,抗番茄黄化曲叶病毒病、烟草花叶病毒病和番茄灰叶斑病等,适宜华北地区早春保护地栽培。     

安徽省农业气象灾害时间分布特征与灰色关联分析

为了探寻安徽省农业气象灾害的分布特点以及各种气象灾害对粮食生产的影响,本研究基于安徽省1992—2012年有关气象灾害数据,对4种主要气象灾害(旱灾、水灾、风雹灾、霜冻灾)的分布特征进行统计分析,并采用灰色关联分析研究其对安徽省粮食单产的影响。结果表明:1992—2012年,安徽省气象灾害具有发生频率高、波动大的特点,但整体上呈下降趋势。旱灾和水灾是发生面积较大的气象灾害,且常在时间上交织、空间上并存。灰色关联分析表明,4种气象灾害对粮食产量影响顺序:风雹灾水灾旱灾霜冻灾,说明风雹灾是影响粮食产量最主要的气象灾害,其次是水灾、旱灾,霜冻灾影响较小。本研究可为安徽省防灾减灾措施的制定提供决策依据。     

基于级联卷积神经网络的番茄花期识别检测方法

对作物花期状态的准确识别是温室作物执行授粉的前提。为提高花期状态识别的准确度，该研究以温室番茄为例提出了一种基于级联卷积神经网络的番茄花朵花期识别方法。首先采用改进的端到端的特征金字塔网络FS-FPN实现番茄花束的分割，然后采用Prim最小生成树对分割后的花束图片进行花期识别优先级排序，最后将已排序的分割花束图片输入改进的Yolov3网络，实现番茄花朵花期状态的精准识别。在由1600幅包含花蕾期、全开期、谢花期、初果期4类花期状态的番茄花束图像构成的数据集上，所提方法对番茄花朵花期平均检测时间为12.54 ms，平均检测精度分别比Mask R-CNN和SPP-Net提高了3.67%和2.39%，识别错误率比改进前的Yolov3网络降低了1.25%。最终将该方法部署到番茄授粉机器人上，并在大型玻璃温室内进行验证，结果表明，所提方法对番茄花朵各花期的检测精度分别为花蕾期85.71%、全开期95.46%、谢花期62.66%、初果期88.34%，该研究结果可为智能授粉机器人的精准作业提供重要依据。