北方地区小西葫芦黄花叶病毒的酶联检测和西瓜品种抗病性鉴定

应用DAS-ELISA对分别于1998年在河南、陕西和1999年在河南、陕西、河北和山西采集的葫芦科病毒病样本84份、186份进行小西葫芦黄花叶病毒的检测,表明该病毒在这些地区广泛发生,ZYMV阳性检出率分别为79.8%和57.5%熏在北京和河南临颖县ZYMV发生率较低熏≤23.1%,在其余地区即河北石家庄,河南郑州、开封、孟津,山西运城,陕西西安其阳性检出率在40.0%～92.9%之间。除冬瓜样本没有ZYMV外,西瓜、甜瓜、南瓜、丝瓜、小西葫芦、苦瓜、黄瓜和瓠子均受到ZYMV的侵染。不同地区、年份和作物之间ZYMV阳性检出率存在差异。对18份西瓜品种的抗ZYMV温室人工接种鉴定的试验表明,目前推广的一些品种对ZYMV缺乏抗病性。     

Comparison of the nucleotide and amino acid sequences of parental and attenuated isolates of Zucchini yellow mosaic virus

To identify possible sites of viral attenuation, the complete nucleotide sequences of two isolates of Zucchini yellow mosaic virus (ZYMV) were determined; a severe isolate Z5-1 and an attenuated isolate from Z5-1 (designated ZYMV-2002). The viral genome of both isolates consisted of 9593 nucleotides in size and contained an open reading frame encoding a single polyprotein of 3080 amino acids. Comparison of the nucleotide sequences for Z5-1 and ZYMV-2002 revealed 14 nucleotide mutations, resulting in seven amino acid substitutions with four in the HC-Pro region, two in the CI region, and one in the NIb region. These results provide a genetic basis for future manipulation of the ZYMV reverse genetics system. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB188115 and AB188116     

西葫芦新品种引种试验

通过对11个西葫芦品种的对比试验,依据产量、品质性状的测定结果以及市场认可度,筛选的优良品种为冬美、京冬玉。     

罗汉果种子传播小西葫芦黄化花叶病毒的检测

为了解小西葫芦黄化花叶病毒（Zucchini yellow mosaicvirus,ZYMV）是否能通过罗汉果种子传播。本研究通过取ZYMV阳性的罹病罗汉果植株种子700粒,400粒直接播种于防虫网室中,300粒经表面消毒处理后播种于MS培养基上。最终获得213棵罗汉果实生苗。对所获得实生苗进行生物学鉴定及ZYMV特异性RT-PCR检测,均未检测到ZYMV阳性植株。结果表明ZYMV不能经罗汉果种子传播,或其传播率很低。     

那氏778诱导剂浸种对西葫芦生长发育及产量品质的影响

以清水浸种为对照,进行了不同浓度那氏778诱导剂浸种试验.结果表明:那氏 778诱导剂浸种处理后西葫芦种子出苗速度变慢,出苗率略有降低;但出苗后,适宜浓度的诱导剂浸种处理其生育进程加快,第一雌花出现和开花时间较早,果实生长速度快且坐果率高,根瓜采收提前1～6 d,第1次采收量和总产量均比对照明显增加,以浓度为1:60的处理增产最为显著,第1次采收量和总产量比对照分别增产78.8%和23.5%.适宜浓度的那氏778诱导剂浸种后畸形果率降低,且有利于改善品质,可溶性糖、维生素C、蛋白质和游离氨基酸含量均比对照有所提高,硝酸盐含量下降.同时,经那氏778诱导剂浸种的处理,抗灰霉病的能力明显提高,灰霉病发生率比对照减少26.9%～62.6%;综合考虑上述诸方面因素,以浸种浓度1:60的处理诱导效果最好.     

螯合剂HIDS对亚适温下西葫芦幼苗的影响

以西葫芦品种‘农园1号’为实验材料,设置7个处理：适温处理（昼/夜25℃/16℃）,浇灌1/3倍山崎黄瓜配方营养液（CK）;亚适温处理（昼/夜16℃/8℃）,浇灌1/3倍山崎黄瓜配方营养液（TCK）;亚适温条件下分别用1/3倍山崎黄瓜配方营养液配成HIDS浓度为100、400、700、1000、1300mg·L−1的营养液浇灌（分别为T1、T2、T3、T4和T5处理）,西葫芦幼苗在人工气候室内培养18d后,分析添加HIDS对西葫芦幼苗生长指标,叶片渗透调节物质、MDA、抗氧化酶活性、光合特性及荧光参数的影响。结果表明：TCK处理西葫芦幼苗生长状况、抗氧化酶活性以及光合能力均比CK处理差。亚适温下不同浓度HIDS处理均可缓解亚适温对西葫芦幼苗生长的抑制,最适浓度为700mg·L−1（T3）,其单株叶面积、全株鲜重、干重、可溶性蛋白、脯氨酸含量、POD、SOD、CAT活性比TCK处理显著提高,MDA含量显著降低;叶绿素总量以及净光合速率、蒸腾速率、胞间二氧化碳浓度和气孔导度显著提高;叶片Fv/Fm、ΦPSⅡ、Fv'/Fm'、ETR、qP升高,NPQ降低,可缓解亚适温对西葫芦光合器官造成的伤害。因此,添加适宜浓度的HIDS可通过促进西葫芦幼苗生物量积累及叶片中渗透调节物质的积累,减轻细胞膜脂过氧化程度,提高抗氧化酶活性和光合能力,以此提高西葫芦幼苗对亚适温的适应能力,以HIDS浓度为700mg·L−1时各指标最接近CK处理,缓解效果最好。     

新疆小西葫芦黄化花叶病毒的分子鉴定与序列分析

在新疆石河子地区采集具有黄化、花叶和疱斑等症状的多种葫芦科作物,分别提取总RNA,利用小西葫芦黄化花叶病毒(Zucchini yellow mosaic virus,ZYMV)特异性引物进行RT-PCR扩增,其中来自黄瓜、甜瓜和丝瓜的模板RNA上可以得到1185bp片段。将扩增得到的片段克隆到pMD18-T上,序列分析结果表明:该片段含有1185个核苷酸,包含完整CP基因。与ZYMV Florida分离物、California、韩国KR-PA分离物、新加坡S及中国北京CH-BJ等GenBank报道的7个不同地域、不同种寄主植物上的分离物序列进行同源性分析的结果显示:ZYMV新疆分离物的CP基因核苷酸与其它11种ZYMV分离物的同源性高达99.42%,氨基酸序列同源性高达98.48%;新疆5个来源于不同寄主植物的ZYMV,不存在明显的遗传变异性,同源性高达97%,而与北京CH-BJ,Reunion和Singapore分离物相比存在较大差异,说明ZYMV地域的相关性要大于寄主上相关性。     

合肥地区发生的西瓜花叶病的病原鉴定

 从合肥市郊区的西瓜病叶上分离出一病毒分离物,该分离物的热钝化温度为60~65℃,稀释终点为10^-4~10^-5,寄主体外存活期为20~23 d,电镜观察病毒粒子约750 nm×13 nm。参考已发表的小西葫芦黄化花叶病毒(Zucchini yellow mosaic virus,ZYMV) CP基因序列设计引物,RT-PCR扩增得到CP基因片段。将该CP片段克隆到pGEM-3Zf (+)中,测序结果表明,该CP基因全长840 nt,共编码279个氨基酸,与国内报道的ZYMV CP基因的核苷酸序列同源性为83.0%~97.3%,氨基酸序列的同源性为91.8%~99.8%,证明该分离物为ZYMV。     

Effectiveness of whey against zucchini squash and cucumber powdery mildew

The effectiveness of whey against powdery mildew (Podosphaera xanthii) of cucumber and zucchini squash was tested in greenhouse experiments. Plants were sprayed once or twice a week with whey at concentrations of 0, 5, 10, 15, 20, 25, and 30% in water. Severity of powdery mildew was estimated weekly by visual assessment of individual leaves and scored as percentage of leaf area affected. Effectiveness of treatments did not differ significantly when applied weekly compared to twice a week. In each instance, powdery mildew severity correlated negatively with whey concentration. For cucumber, the rate of the disease progress in the control ranged from 0.45 to 0.75. Disease progressed more slowly in plants treated with 25–30% whey than when lower concentrations were used. The rate of disease progress varied from 0.12 to 0.33 in plants treated once a week and from 0.13 to 0.17 when applied twice a week. Similar tendencies were observed for zucchini, but disease progressed more rapidly and the final disease severity was higher than in cucumber. Effectiveness of whey applied twice a week at concentrations of 15–30% did not differ significantly among treatments; in these treatments the rate of disease progress was about 0.23 and significantly higher than the value for the water controls (about 0.80). Plants treated with 30% whey often exhibited symptoms of phytotoxicity. The data indicated that whey effectively controlled powdery mildew in cucumber and zucchini. Further studies are needed to optimize the concentration and timing of whey applications for mildew management in commercial crops.     

小西葫芦黄花叶病毒山东南瓜和丝瓜分离物全基因组序列分析

在山东泰安采集到两个表现黄花叶症状的丝瓜和南瓜样品中检测到小西葫芦黄花叶病毒(Zucchini yellow mosaic virus,ZYMV);通过RT-PCR和RACE方法获得了两个分离物的全基因组序列,分别命名为CN:Cm:17(丝瓜分离物)和CN:Lc:17(南瓜分离物)。结果表明,CN:Cm:17和CN:Lc:17基因组除Poly(A)尾巴外,分别含有9 593和9 591个核苷酸(Nucleotides,nt),通过多聚蛋白策略编码一个含3080个氨基酸的多聚蛋白。CN:Cm:17和CN:Lc:17在氨基酸水平上一致率为96.7%;CN:Cm:17与韩国分离物KR:RDA:07的氨基酸一致率最高(97.8%); CN:Lc:17与CN:CJLX30535:14和CN:spider131932:13的一致率最高(98.4%)。重组分析发现,CN:Lc:17具有显著重组事件,为KR:KR-PA:05和CN:Cm:17的重组体。基于多聚蛋白编码序列的系统进化聚类结果显示分组与分离物的地理起源存在密切相关性,所有中国分离物分别聚集在A-Ⅴ和A-Ⅵ亚组。