Aphid-borne Zucchini yellow mosaic virus （ZYMV） is one of the most economically important viruses of cucurbitaceous plants. To survey and control this virus, it is necessary to develop an efficient detection techniq...Aphid-borne Zucchini yellow mosaic virus （ZYMV） is one of the most economically important viruses of cucurbitaceous plants. To survey and control this virus, it is necessary to develop an efficient detection technique. Using purified ZYMV virion and the conventional hybridoma technology, three hybridoma cell lines （16A11, 5A7 and 3B8） secreting monoclonal antibodies （MAbs） against ZYMV Zhejiang isolate were obtained. The working titers of the ascitic fluids secreted by the three hybridoma cell lines were up to 10^-7 by indirect enzyme-linked immunosorbent assay （ELISA）. All MAbs were isotyped as IgG1, kappa light chain. Western blot analysis indicated that the MAb 3B8 could specifically react with the coat protein of ZYMV while MAbs 5A7 and 16A11 reacted strongly with a protein of approximately 51 kDa from the ZYMV-infected leaf tissues. According to this molecular weight, we consider this reactive protein As likely to be the HC-Pro protein. Using these three MAbs, we have now developed five detection assays, i.e., antigen-coated-plate ELISA （ACP-ELISA）, dot-ELISA, tissue blot-ELISA, double-antibody sandwich ELISA （DAS-ELISA）, and immunocapture-RT-PCR （IC-RT-PCR）, for the sensitive, specific, and easy detection of ZYMV. The sensitivity test revealed that ZYMV could be readily detected respectively by ACP-ELISA, dot-ELISA, DAS-ELISA and IC-RT-PCR in 1：163840, 1：2560, 1：327680 and 1：1 310720 （w/v, g mL-1） diluted crude extracts from the ZYMV-infected plants. We demonstrated in this study that the dot-ELISA could also be used to detect ZYMV in individual viruliferous aphids. A total of 275 cucurbitaceous plant samples collected from the Zhejiang, Jiangsu, Shandong and Hainan provinces, China, were screened for the presence of ZYMV with the described assays. Our results showed that 163 of the 275 samples （59%） were infected with ZYMV. This finding indicates that ZYMV As now widely present in cucurbitaceous crops in China. RT-PCR followed by DNA sequencing and sequence analy展开更多
采用RT-PCR和RACE-PCR的方法从西葫芦果实中克隆了ABA生物合成关键酶NCED基因保守片段及其3'端,命名为CpNCED1,并通过Real time RT-PCR方法分析了CpNCED1基因在果实发育和成熟过程中的表达。结果表明:CpNCED1基因的DNA序列及其推导...采用RT-PCR和RACE-PCR的方法从西葫芦果实中克隆了ABA生物合成关键酶NCED基因保守片段及其3'端,命名为CpNCED1,并通过Real time RT-PCR方法分析了CpNCED1基因在果实发育和成熟过程中的表达。结果表明:CpNCED1基因的DNA序列及其推导的蛋白质序列与番茄、鳄梨及柑橘果实相同基因的同源性分别为61.90%(LeNCED1)、62.15%(PaNCED1)、62.55%(PaNCED3)、54.67%(CsNCED1)、56.23%(CsNCED2)和69.75%、65.74%、66.67%、68.75%、71.76%。果肉中CpNCED1基因表达共出现了3个高峰,分别在花后5 d,花后20 d及花后35 d,并分别与ABA积累相对应;果皮中CpNCED1表达在花后25 d达到最大值,而瓤和种子中CpNCED1的表达没有显著变化。果皮和种子中ABA含量与其CpNCED1的表达基本一致。西葫芦果实在生长发育过程中乙烯产生量很低,为非跃变型果实;呼吸高峰出现在花后20 d。上述结果说明,西葫芦果实的成熟主要和ABA调控有关,而乙烯对果实成熟的贡献很小。展开更多
Plants respond to drought stress with different physical manners, such as morphology and color of leaves. Thus, plants can be considered as a sort of living-sensors for monitoring dynamic of soil water content or the ...Plants respond to drought stress with different physical manners, such as morphology and color of leaves. Thus, plants can be considered as a sort of living-sensors for monitoring dynamic of soil water content or the stored water in plant body. Because of difficulty to identify the early wilting symptom of plants from the results in 2D （two-dimension） space, this paper presented a preliminary study with 3D （three-dimension）-based image, in which a laser scanner was used for achieving the morphological information of zucchini （Cucurbita pepo） leaves. Moreover, a leaf wilting index （DLWIF） was defined by fractal dimension. The experiment consisted of phase-1 for observing the temporal variation of DLWIF and phase-2 for the validation of this index. During the experiment, air temperature, luminous intensity, and volumetric soil water contents （VSWC） were simultaneously recorded over time. The results of both phases fitted the bisector （line： 1：1） with R2=0.903 and REMS=0.155. More significantly, the influence of VSWC with three levels （0.22, 0.30, and 0.36 cm3 cm-3） on the response of plant samples to drought stress was observed from separated traces of DLWIF. In brief, two conclusions have been made： （i） the laser scanner is an effective tool for the non-contact detection of morphological wilting of plants, and （ii） defined DLWIF can be a promising indicator for a category of plants like zucchini.展开更多
基金supported by the National Natural Science Foundation of China(31272015)the National Basic Research Program(973)of China(2014CB138400)the Special Fund for Agro-scientific Research in the Public Interest,China(201303021,201303028)
文摘Aphid-borne Zucchini yellow mosaic virus （ZYMV） is one of the most economically important viruses of cucurbitaceous plants. To survey and control this virus, it is necessary to develop an efficient detection technique. Using purified ZYMV virion and the conventional hybridoma technology, three hybridoma cell lines （16A11, 5A7 and 3B8） secreting monoclonal antibodies （MAbs） against ZYMV Zhejiang isolate were obtained. The working titers of the ascitic fluids secreted by the three hybridoma cell lines were up to 10^-7 by indirect enzyme-linked immunosorbent assay （ELISA）. All MAbs were isotyped as IgG1, kappa light chain. Western blot analysis indicated that the MAb 3B8 could specifically react with the coat protein of ZYMV while MAbs 5A7 and 16A11 reacted strongly with a protein of approximately 51 kDa from the ZYMV-infected leaf tissues. According to this molecular weight, we consider this reactive protein As likely to be the HC-Pro protein. Using these three MAbs, we have now developed five detection assays, i.e., antigen-coated-plate ELISA （ACP-ELISA）, dot-ELISA, tissue blot-ELISA, double-antibody sandwich ELISA （DAS-ELISA）, and immunocapture-RT-PCR （IC-RT-PCR）, for the sensitive, specific, and easy detection of ZYMV. The sensitivity test revealed that ZYMV could be readily detected respectively by ACP-ELISA, dot-ELISA, DAS-ELISA and IC-RT-PCR in 1：163840, 1：2560, 1：327680 and 1：1 310720 （w/v, g mL-1） diluted crude extracts from the ZYMV-infected plants. We demonstrated in this study that the dot-ELISA could also be used to detect ZYMV in individual viruliferous aphids. A total of 275 cucurbitaceous plant samples collected from the Zhejiang, Jiangsu, Shandong and Hainan provinces, China, were screened for the presence of ZYMV with the described assays. Our results showed that 163 of the 275 samples （59%） were infected with ZYMV. This finding indicates that ZYMV As now widely present in cucurbitaceous crops in China. RT-PCR followed by DNA sequencing and sequence analy
基金the Chinese-German Center for Scientific Promotion (Chinesisch-Deutsches Zentrum für Wissenschaftsfrderung) under the Project of Sino-German Research Group (GZ494)the Beijing Municipal Education Commission for Building Scientific Research and Scientific Research Base (2008BJKY01)+1 种基金the German Academic Exchange Service (DAAD),and China Scholarship Council (CSC) for enhancing our cooperationthe International Cooperation Fund of Ministry of Science and Technology, China (2010DFA34670)
文摘Plants respond to drought stress with different physical manners, such as morphology and color of leaves. Thus, plants can be considered as a sort of living-sensors for monitoring dynamic of soil water content or the stored water in plant body. Because of difficulty to identify the early wilting symptom of plants from the results in 2D （two-dimension） space, this paper presented a preliminary study with 3D （three-dimension）-based image, in which a laser scanner was used for achieving the morphological information of zucchini （Cucurbita pepo） leaves. Moreover, a leaf wilting index （DLWIF） was defined by fractal dimension. The experiment consisted of phase-1 for observing the temporal variation of DLWIF and phase-2 for the validation of this index. During the experiment, air temperature, luminous intensity, and volumetric soil water contents （VSWC） were simultaneously recorded over time. The results of both phases fitted the bisector （line： 1：1） with R2=0.903 and REMS=0.155. More significantly, the influence of VSWC with three levels （0.22, 0.30, and 0.36 cm3 cm-3） on the response of plant samples to drought stress was observed from separated traces of DLWIF. In brief, two conclusions have been made： （i） the laser scanner is an effective tool for the non-contact detection of morphological wilting of plants, and （ii） defined DLWIF can be a promising indicator for a category of plants like zucchini.
