不同温度下的豌豆蚜实验种群生命表研究

在6种温度(12、15、18、21、24℃和27℃)和相对湿度为80%的条件下,研究了温度对豌豆蚜发育和增殖的影响。组建了生命表,并统计出种群内禀增长率、净增殖率、世代平均周期、种群加倍时间和周限增长率等种群动态参数。结果表明,在试验温度范围内,豌豆蚜的发育历期随温度升高而缩短。种群的内禀增长率在24℃条件下最大(rm=0.281 0),相应的种群净增殖率(R0)、周限增长率(λ)、平均世代周期(T)和种群加倍时间(t)分别为17.030 9、1.324 5、10.089 3 d和2.466 7 d。     

黄土丘陵区垄沟改良措施对玉米水分利用效率的影响

选择延安地区作为黄土丘陵区的代表,在延安地区应用地膜、液态地膜、保水剂、PAM(土壤改良剂)和间作套种措施优化大垄沟种植方式种植玉米,分析了不同优化措施种植玉米的水分利用效率。结果表明,地膜、保水剂、PAM和间作方式有利于大垄沟较好地保持土壤水分,其保水能力分别比平种高出17.4%、8.5%、11.6%、5.3%;液态地膜、地膜、间作和PAM表现出很好的增产效果,其产量分别比平种高出77.7%、72.0%、64.1%、30.8%。     

A蛋白斑点免疫金染色和双抗体夹心酶联检测齿兰环斑病毒的比较

A蛋白斑点免疫金染色检测齿兰环斑病毒提纯病毒的灵敏度为1.56 ng/μl,检测病汁液的稀释倍数为640倍.同时采用碱性磷酸酯酶标记抗体IgG.IgG包被酶联板的浓度为1 μg/ml,酶标抗体以1 /1000稀释使用.DAS-ELISA技术检测ORSV提纯病毒的灵敏度为0.048 ng/μl,检测病汁液的稀释倍数为20480倍.     

Identification of divergent variants of Grapevine virus A

Eight isolates of Grapevine virus A (GVA), which induced different symptoms in leaves of Nicotiana benthamiana, were recovered from various grapevines. The dsRNA patterns of two isolates, which consistently induced mild vein clearing (referred here as mild isolates of GVA) were similar, but different from those of other isolates of GVA. Analysis based on overall nucleotide (nt) sequence identity in the 3 terminal part of the GVA genome, comprising part of ORF3 (putative movement protein, MP), entire ORF4 (capsid protein, CP), entire ORF5 and part of 3 UTR, revealed that GVA isolates separate into three groups (I, II, III), sharing 91.0–99.8% nt sequence identity within groups and 78.0–89.3% nt sequence identity between groups. Mild isolates of the virus were group III and shared only 78.0–79.6% nt sequence identity with the other isolates. The comparison of predicted amino acid sequences for MP and CP revealed many amino acid alterations, revealing distinct local net charges of these proteins for mild isolates of the virus. Based on both conserved and divergent nt regions in the CP and ORF5, oligonucleotide primers were designed for the simultaneous RT-PCR detection of all GVA isolates and for the specific detection of the most divergent virus variants represented here by mild isolates of the virus.     

2-硝基-1-芳乙烯类化合物的设计合成及其对稻瘟病菌的抑制活性

为了筛选得到对稻瘟病菌Magnaporthe grisea具有较高抑菌活性的新型化合物,根据稻瘟病菌中1,3,8-三羟基萘酚还原酶（3HNR）的结构信息,设计合成了系列2-硝基-1-芳乙烯（2a~2e）和2-溴-2-硝基-1-芳乙烯（3a,3b）目标化合物,并测试了其对3HNR和稻瘟病菌的抑制活性,同时运用分子对接方法对目标化合物与3HNR可能的结合模式进行了分析。结果表明:大部分目标化合物对3HNR都有很好的抑制作用（IC503的抑制活性最好,IC50值为0.53 μmol/L。在50 μg/mL下,目标化合物对稻瘟病菌的生长具有不同程度的抑制作用,其中2e、3a和3b的抑制率高于96%;3a和3b的EC50值分别为16.4和11.6 μg/mL。分子对接方法分析结果表明,硝基苯乙烯骨架结构与稻瘟病菌的3HNR活性空腔的氨基酸残基有较好的相互作用,其中化合物3中的溴原子可与3HNR中Tyr223和Tyr178的羟基形成氢键,从而解释了化合物3对3HNR有较好抑制作用的原因。     

灰飞虱体内水稻条纹病毒的检测

为评价灰飞虱体内水稻条纹病毒检测方法的适用范围,利用已经制备的水稻条纹病毒(Rice stripe virus,RSV)的多克隆抗体SV21和单克隆抗体3B9,采用多孔板间接ELISA、DIBA和Western blotting等三种方法进行单头灰飞虱Laodelphax striatellus Fallén(SBPH)体内RSV检测。结果表明,检测灵敏度以多孔板间接ELISA最高,其次为DIBA,Western blotting最低;用单克隆抗体3B9检测灵敏度高于多克隆抗体SV21。RT-PCR、IC-RT-PCR和DB-RT-PCR三种方法中,RT-PCR对单头灰飞虱稀释到400倍可检测到病毒;IC-RT-PCR在多克隆抗体SV21稀释浓度大于500倍时检测不出RSV,单克隆抗体3B9稀释浓度大于800倍时检测不到RSV;DB-RT-PCR检测结果显示,单头灰飞虱在稀释400倍后均无阳性反应。     

基于全基因序列的中国北方3省(区)马铃薯Y病毒遗传多样性分析

本研究以马铃薯Y病毒(PVY)全基因组为基础,分析吉林、黑龙江和内蒙古3省(区)PVY群体遗传多样性和群体分化,并评估突变、重组、选择等遗传力所起的作用。根据已报道的PVY全基因序列保守区设计4对引物,采用片段重叠法对来自内蒙古和吉林的24个PVY分离物全基因序列进行测定,并联合NCBI中已登录的9个黑龙江分离物全基因组序列进行遗传多样性参数评估、群体分化检验和分子变异等分析。结果显示,我国北方3省(区)PVY群体遗传多样性高,其中内蒙古和黑龙江PVY群体遗传多样性高于吉林群体,并且3个群体之间呈现一定程度的遗传分化。分子变异分析发现在PVY基因组中存在1 786个变异位点,表明我国北方3省(区)PVY群体变异程度较高,并且这种高变异度有85.54%来自各个马铃薯种植区内PVY个体的遗传变异。重组分析和系统发育分析发现,我国北方3省(区)PVY群体中重组株系占比高达90.3%,并具有明显的株系多样性,表明PVY重组株系已成为我国北方3省(区)马铃薯种植区的流行株系。选择压力分析显示,使用FEL和IFEL法分别检测出501个和315个净化压力选择位点,这表明3省(区)PVY群体受净化选择压力为主。以上结果表明,中国北方3省(区)PVY群体遗传多样性高,突变、重组和自然选择都对遗传多样性和群体分化存在一定影响。     

玉米抗粗缩病遗传及分子标记研究

Maize rough dwarf disease caused by Rice black-streaked dwarf virus (RBSDV) is transmitted by planthopper in China. Identification and development of resistant hybrids are complicated because of the inconsistencies in viral disease pressure every year. Marker-assisted selection can provide means for main-taining virus resistance alleles even in the absence of disease. In this paper a F₂ segregation population was constructed to identity the molecular markers linked to the resistance gene using a cross between a resistant and a susceptible parents (Qi319×Ye107). Fifteen-day-old seedlings of F₂ population were exposed to small brown planthoppers carrying RBSDV for 3 days in specific inoculation chamber. The inoculated plants were transplanted to screenhouse after removing the insects completely. In plant maturity stage the disease resistance of all the individuals were visually assessed. The results showed that 17, 8, 11, 51 and 122 plants were scaled from 0-4 respectively, in which 0 means no symptoms and 4 represents highly susceptible. Chi-square test demonstrated that the segregation ratio of phenotype was 1∶15 (resistant: susceptible) or 1∶6∶9 (resistant∶moderate∶susceptible) in the F₂ population, indicating RBSDV resistance of maize was controlled by two recessive genes. The F₂ individuals DNA were extracted and 261 SSR (simple sequence repeat) primers derived from maize genome ten chromosomes were selected from maize GDB database to construct genetic linkage map. The linkage map consisted of 71 polymorphic SSR markers, spanning a genetic distance of 996.6 cM with an average interval of 14.0 cM between adjacent markers. The resistant and susceptible gene pools were set up for BSA (bulked segregant analysis) and 6 polymorphism markers were obtained with BSA-SSR method between the two pools. The F₂individuals were further analyzed with 6 polymorphism markers. Chi-square test showed that phi 051, umc1407 and umc1432, mapped on chromosome 7 and 10, exhibited segregation distortion significantly and very significantly in susceptible individuals. These three SSR markers were identified as potential markers linked to the resistant loci.     

引起浙贝母黑斑病的交链格孢单抗及检测试纸条的制备

浙贝母黑斑病是危害浙贝母茎叶的重要真菌性病害之一, 该病侵染叶片产生叶斑?叶枯, 严重时全株枯死?本研究制备了针对浙贝母黑斑病致病菌交链格孢Alternaria alternata的单克隆抗体(单抗), 并开发了胶体碳免疫层析试纸条用于黑斑病早期精准检测?以交链格孢提取液免疫Bal b/c小鼠, 采用杂交瘤技术制备单抗, 间接ELISA法和Western blot分析单抗的效价?灵敏度?特异性?抗体类型及结合蛋白?以双抗夹心法制备胶体碳免疫层析试纸条?得到3种针对交链格孢的单抗及其杂交瘤细胞株AaA1?AaC2和AaC5?3种单抗对交链格孢均具有高度特异性, 灵敏度为12.21~24.41 ng/mL, 效价≥6.40×105, 抗体类型分别为IgG2a?IgG3和IgG3, 结合蛋白的分子量分别为37?62 kDa和66 kDa?以AaA1为碳标单抗, AaC2为划膜单抗的胶体碳试纸条检测交链格孢的方法快速简便?准确灵敏?经济环保?从田间采样到出结果用时少于15 min, 样品简单研磨后取汁液加入加样孔即完成全部操作, 对交链格孢抗原的检测灵敏度为6.25 μg/mL, 试纸条采用环保材料制备, 单个试纸条的成本约1.7元?     

用免疫分离法检测稻种上的白叶枯病菌

 免疫分离法是一种新型的、高度灵敏和准确的种菌带菌检验法,它结合了利用专化性抗血清的选择法吸附作用和活细菌可形成菌落的两方面的优点,从种苗材料中分离到目标细菌,用纯菌测试它的回收率为50~70%,最低检出率为50~100个菌/ml,人工喷接种和病田收获的病种带菌率达87%,平均每粒自然病种上的细菌数在100个左右。免疫分离所得菌落经致病性测定,大多数为白叶枯菌。