冬瓜杂种种子纯度鉴定的SSR分析

种子纯度是种子质量的核心指标,目前冬瓜、节瓜杂种优势已被广泛地应用于生产。探索出一种快速、准确的冬瓜、节瓜品种纯度鉴定方法是确保种子质量的技术保证,也是品种纯度检测实现商业化的前提。本研究以‘绿优’冬瓜和‘梅花8号’节瓜及其各自亲本为试验材料,利用SSR分子标记技术对亲本及杂种之间的多态性进行分析。结果显示,从200对SSR引物中筛选出2对多态性引物scaffold4775和scaffold5674,用于‘绿优’冬瓜和‘梅花8号’节瓜杂种种子纯度的鉴定。其扩增片段电泳条带清晰可辨,杂交种中呈现双亲的互补带型,SSR标记表现出稳定的显性和共显性,能将‘绿优’冬瓜和‘梅花8号’节瓜与其父母本区分开来。分子检测平均纯度均为98%,田间纯度鉴定分别为98%和99%,结果基本一致,表明引物scaffold4775和scaffold5674可用于‘绿优’冬瓜和‘梅花8号’节瓜的纯度鉴定。SSR分子标记呈现出经济快速、准确等特点,在冬瓜、节瓜品种纯度鉴定上其应用前景十分广泛。     

基于分子生态学网络分析松嫩退化草地土壤微生物群落对施氮的响应

【目的】 氮素输入影响着全球草地生态系统的可持续性,关注施氮对土壤微生物群落的影响及其分子生态网络,为草地退化修复提供理论依据。【方法】 以松嫩退化羊草草地为研究对象,通过施氮和未施氮处理,利用高通量测序和随机矩阵网络构建理论构建土壤微生物群落分子生态网络。探讨氮素管理对退化羊草草地土壤微生物群落结构及网络的影响,氮添加条件下微生物网络结构中的关键微生物变化规律,以及该过程中微生物之间的互作关系,解析外源氮素添加条件下土壤细菌动态变化的关键结点和规律。【结果】 在门分类水平上施氮处理草地有细菌门22个,未施氮处理23个。7个菌门是施氮和未施氮处理草地的优势菌门,其中变形菌门（Proteobacteria）是含有OTU数量最多的门类,约占总序列的30.46%,酸杆菌门（Acidobacteria）是含有OTU数量次之的门类,约占总序列的30.15%,芽单胞菌门（Gemmatimonadetes）是含有OTU数量第3的门类,约占总序列的8.14%,放线菌（Actinomycete）约占总序列的6.15%,绿弯菌门（Chloroflexi）、拟杆菌门（Bacteroidetes）和硝化螺旋菌门（Nitrospirae）三者约占总序列的17.16%。施氮处理草地土壤微生物中的变形菌门、放线菌门、拟杆菌门的相对丰度均显著高于未施氮处理草地土壤（P<0.01）;未施氮草地土壤中绿弯菌门、酸杆菌门、芽单胞菌门相对丰度显著高于施氮草地土壤（P<0.01）,其他各门细菌施氮与未施氮处理间未呈现出显著差异。表征网络的正向连接比、平均路径长度、平均聚类系数和模块性均为施氮处理显著低于未施氮处理（P<0.001）。在土壤的分子生态网络中,未施氮处理有16个模块枢纽（Zi>2.5,Pi≤0.62）,施氮处理有6个模块枢纽,均属于酸杆菌门、芽单胞菌门和放线菌门。施氮导致土壤微生物种间关系改变,进而改变土壤整体生态网络。【结论】 施氮降低了退化草地土壤网络结构的复杂程度和紧密性;降低了退化草地土壤中的酸杆菌门和绿弯菌门相对丰度,提高了变形菌门、放线菌门和芽单胞菌门相对丰度。土壤中微生物关键物种（OTU）由16个（未施氮）减少为6个,且二者土壤中均没有重叠OTU,这表明施氮调控微生物群落网络的关键物种,进而改变其分子生态网络结构。     

A data transfer method for improving seed identification of maize (Zea mays) haploid breeding based on genetic similarity

Maize haploid breeding technology is able to identify haploid seeds non‐destructively, rapidly and at low cost with the help of Near‐infrared (NIR) spectral analysis. However, due to the hybridization of numerous parents and the low production rate of haploid, the haploid data collection becomes a burden for engineering this technology. Biologically, there are considerable similarities between the progeny of the same female parent and different male parents. Based on this advantage, similar spectral data can be transferred when the NIR technology is employed. A revised method of Transfer adaptive boost (TrAdaBoost) is proposed to improve identifying for the backpropagation neural network (BPNN) classifier. To avoid the negative transfer, a screening thresh is used to select out similar data, and the amount of these data are limited to join current training. The results show that the identification performances are improved significantly when the data amount is small. This method shows a high ability to make the seed identification more convenient for engineering maize haploid breeding.     

中华稻蝗Knickkopf(Knk)基因分子特性和生物学功能

基于中华稻蝗转录组数据库,采用生物信息学方法搜索获得1条OcKnk基因全长cDNA序列,采用qRT-PCR检测其组织部位表达特性和其在表皮发育过程中表达情况,明确其分子特性,采用RNAi技术结合表型观察,研究其对中华稻蝗蜕皮和生长发育的影响,以明确其生物学功能。组织部位表达结果显示其在中华稻蝗体壁、前肠和脂肪体表达最高,发育表达结果显示其在表皮不同发育日龄均有表达,且在蜕皮前期和后期表达显著高于其他日龄。生物学功能研究表明,注射dsRNA后,多数虫体难以成功蜕去旧表皮,导致死亡,少部分可蜕至下一龄期,但活动力较低,行动缓慢,最终死亡。研究结果表明OcKnk参与昆虫生长发育和蜕皮过程,可作为重要靶标基因,为下一步害虫防治提供理论基础。     

四川地区猕猴桃病毒1的RT-PCR检测及外壳蛋白基因序列分析

为了明确近期新西兰报道的侵染猕猴桃的新病毒——猕猴桃病毒1（Actinidia virus 1，AcV-1）在四川地区猕猴桃上的发生情况及其分子特性，采用RT-PCR对来自四川6个地区疑似感染病毒的90份猕猴桃主栽品种‘红阳’和‘金果’的叶片样品进行检测。结果表明，22份样品为AcV-1阳性，检出率为24.4%。将获得的5个AcV-1分离物和已报道的新西兰分离物K75的外壳蛋白（coat protein，CP）基因序列进行比对，结果表明，分离物间核苷酸序列和氨基酸序列的相似性分别为84.8% ~ 97.1%和89.7% ~ 99.6%，其中除邛崃分离物HYH5与新西兰分离物K75的核苷酸序列相似性较高（97.1%）外，其余4个分离物均低于91%。系统进化分析结果显示，这些分离物主要聚集在3个分支上，分离物HYH5与新西兰分离物K75位于同一分支，其余分离物位于另外2个分支。     

番茄斑萎病毒病抗性种质筛选及人工接种鉴定方法优化

对154份番茄材料(包括普通番茄76份,樱桃番茄78份)进行两年两次田间番茄斑萎病毒病的病情指数调查,筛选出14份对番茄斑萎病毒病具有稳定抗性的番茄材料,利用sw-5-2共显性SCAR标记对田间表现抗病的材料进行分子标记鉴定,发现3份抗病材料携带抗番茄斑萎病毒病的Sw-5基因。为缩短番茄斑萎病毒病人工接种鉴定周期,以含有sw-5的抗病材料H8和感病材料M82为研究对象,设置4、6、8、10片真叶4个接种时期,分别在接种后14、21、28 d进行病情指数调查和抗性分级,结果表明,6片真叶期接种,接种后28 d进行病情调查即可有效鉴别植株番茄斑萎病毒病抗性,与8、10片真叶期接种效果相同,人工接种抗病性鉴定效率显著提升。     

Longitudinal identification of Enterocytozoon bieneusi in dairy calves on a farm in Southern Xinjiang,China

Enterocytozoon bieneusi is the most common species responsible for human and animals microsporidiasis. A total of 250 samples were collected weekly from 25 newborn dairy calves of a farm in Southern Xinjiang, China at one to ten weeks of age. Enterocytozoon bieneusi was identified and genotyped by nested PCR amplification and sequencing of internal transcribed spacer (ITS) region.The cumulative prevalence of E. bieneusi infection was 100% (25/25), and the average infection was 52.0% (130/250). The highest infection rate was recorded at six weeks of age (92.0%, 23/25), and no infection was observed at one and two weeks of age. Sequencing analysis showed nine E. bieneusi genotypes (J, EbpC, PigEBITS5, CHV4, CHC3, CS-9, KIN-1, CH5, and CAM5) were identified. The highest genetic polymorphism was observed at ten weeks of age. Genotype J was the predominant E. bieneusi genotype. Phylogenetic analysis clustered genotype J into Group 2 and other eight genotypes (EbpC, PigEBITS5, CHV4, CHC3, CS-9, KIN-1, CH5, and CAM5), detected in 22 (16.9%, 22/130) samples, into Group 1. Among the genotypes, EbpC, KIN-1, and J have been identified in humans. The highest E. bieneusi infection rate (57.9%, 124/214) was observed in fecal samples with formed feces with no diarrhea (p < 0.01), and high genetic polymorphism was observed in class I fecal samples. The presence of zoonotic E. bieneusi genotypes in dairy calves suggests the possibility of transmitting zoonotic infections to humans. It provides the basic data on dynamic change of E. bieneusi in calves.     

Comparative analysis of genetic effects of wheat‐Dasypyrum villosum translocations T6V#2S·6AL and T6V#4S·6DL

Wheat‐Dasypyrum villosum translocations T6V#2S·6AL and T6V#4S·6DL, carriers of Pm21 and PmV, respectively, confer high resistance to wheat powdery mildew. For better understanding of the difference in genetic effect between them, a RIL population was constructed based on the cross between “Yangmai 18” carrying T6V#2S·6AL and “Yangmai 22” carrying T6V#4S·6DL. Analysis of distribution of the translocations showed that T6V#2S·6AL is much more transmittable than T6V#4S·6DL. By comparing their effects on main agronomic traits, we firstly found that T6V#2S·6AL contributes greatly to top spikelet fecundity, but causes a decrease of 6.7%–10.5% of spike number. No stable effects of T6V#4S·6DL on agronomic traits were found, except for positive effect on plant height. Excitingly, a new recombinant, T6V#4S‐6V#2S·6AL carrying PmV, was screened and proved to have a higher transmission rate than the original translocation T6V#4S·6DL, which will greatly promote the utilization of PmV. The above conclusions of this research will provide important guidance for utilization of Pm21 and PmV more effectively, in wheat powdery mildew resistance breeding.     

70份国外小麦品种(系)的苗期和成株期抗叶锈病鉴定

小麦叶锈病是小麦生产中的重要病害之一,培育持久抗病品种是最经济、有效和环保的方法。本研究用19个不同毒力的叶锈菌小种苗期接种70份国外引进小麦品种(系)及36个已知抗叶锈病基因的载体品种进行抗性鉴定,同时在2016—2017年度分别于河北保定和河南周口对70份国外引进品种进行田间抗叶锈性鉴定。为进一步检测材料中所携带的苗期和成株抗叶锈病基因,利用12个与已知基因紧密连锁的分子标记进行检测,综合基因推导、系谱分析和分子标记检测的结果,在33份材料中鉴定出15个抗叶锈病基因,包括Lr1、Lr2a、Lr26、Lr3ka、Lr11、Lr17、Lr30、Lr10、Lr14a、Lr2b、Lr13、Lr15、Lr21、Lr44和Lr45,田间鉴定筛选出39份品种表现慢锈性。苗期和田间表现表明,国外品种中含有丰富的对我国叶锈菌小种有效的苗期和成株期抗叶锈病基因,可作为小麦抗叶锈病抗源在抗病育种中加以利用。     

星油藤青枯病病原菌鉴定

 星油藤(Plukenetia volubilis L.) 是一种重要的藤本油料植物,在我国华南地区广泛种植。青枯病是近两年在海南星油藤种植区发生的新病害,为探究星油藤青枯病菌的基本特性及种下分化情况,本研究对分离的6株代表菌株进行了相关分析。细菌学鉴定及致病性测定结果表明,该病害是由类茄科雷尔氏菌(Ralstonia pseudosolanacearum)侵染引起。同时,从传统分类及分子生物学不同层面分析了星油藤青枯病菌的遗传分化情况。生理小种及生化变种的测试结果表明,星油藤青枯病菌属于1号生理小种和生化变种Ⅲ;16S rDNA和egl基因部分序列聚类分析显示,星油藤青枯病菌属类茄科雷尔氏菌演化型Ⅰ即亚洲分支菌株,序列变种34。