1株H1N1亚型猪流感病毒的全基因组特征及其对小鼠的致病性

旨在了解河南省猪流感病毒的流行情况及其遗传进化和基因组特征。2018年4月，从河南省某一出现疑似流感症状猪群中采集鼻拭子样品150份用于分离病毒，对分离病毒的全基因组进行序列测定和分析。同时感染6周龄BALB/c小鼠，研究其对小鼠的致病性。结果显示，获得1株H1N1亚型病毒[命名为A/swine/Henan/NY20/2018（H1N1）]。遗传进化表明，其HA和NA基因属于欧亚类禽H1N1分支，PB2、PB1、PA、NP和M基因属于2009甲型H1N1分支，NS基因属于经典H1N1分支。HA蛋白的裂解位点序列为PSIQSR↓GL，具有低致病性流感病毒的分子特征，在小鼠肺和鼻甲有效复制并能引起肺组织病理学变化。本研究分离到1株3源重排H1N1亚型病毒，对小鼠呈现一定致病力，提示应进一步加强对SIV的监测。     

大豆质核互作雄性不育系NJCMS1A与其保持系NJCMS1B的生理生化特性比较分析

以大豆质核互作雄性不育系NJCMS1A及其保持系NJCMS1B的花芽为试验材料,利用试剂盒法和紫外分光光度计测定分析了糖分含量(可溶性糖和淀粉)和总ATPase、蔗糖磷酸合成酶(SPS)、磷酸烯醇式丙酮酸羧激酶(PEPCK)、抗坏血酸氧化酶(AO)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)以及过氧化物酶(POD)等酶活性的变化情况。结果表明:相较于保持系NJCMS1B,总ATPase、PEPCK、AO和CAT活性在不育系NJCMS1A中显著下降;糖分含量、SPS和SOD活性在不育系NJCMS1A中下降水平不显著;POD活性在不育系NJCMS1A中显著上升。根据结果分析推测,与能量代谢或胁迫响应等相关的物质或酶活性在不育系NJCMS1A中的亏损现象可能是NJCMS1A花粉败育的主要原因。     

米槠人工林土壤微生物群落组成对凋落物输入的响应

全球气候变化显著影响森林凋落物数量,进而会对土壤微生物群落造成影响。本研究以亚热带米槠人工林为研究对象,探究不同凋落物量输入处理(凋落物去除、凋落物加倍、对照)下,森林土壤微生物群落组成的变化。结果表明:与去除凋落物相比,凋落物加倍后0~10 cm土壤铵态氮(NH4^+-N)、硝态氮(NO3^--N)、全氮(TN)、有效磷(AP)含量分别显著增加了30.30%、49.66%、12.77%和13.90%。与对照相比,凋落物加倍与去除处理土壤微生物生物量碳(MBC)和氮(MBN)含量分别显著增加和下降(P<0.05),但凋落物加倍与去除处理间无显著差异。凋落物加倍处理下土壤丛枝菌根真菌(AMF)、革兰氏阳性菌[G(+)]、革兰氏阴性菌[G(-)]、放线菌(ACT)、真菌(F)丰度和总磷脂脂肪酸(TPLFA)含量分别比去除凋落物处理的土壤高68.35%、63.35%、82.65%、69.02%、40.56%和65.85%,而土壤革兰氏阳性菌与阴性菌比值、真菌与细菌比值则分别降低11.64%和26.67%。冗余度分析表明,铵态氮是影响该人工林土壤微生物群落组成的最主要环境因子。可见凋落物输入量变化改变了土壤养分有效性,进而显著影响了土壤微生物群落组成,这对进一步深入探究全球气候变化对亚热带森林土壤养分循环的影响具有重要意义。     

Development of a core set of KASP markers for assaying genetic diversity in Brassica rapa subsp. chinensis Makino

Single‐nucleotide polymorphisms (SNPs) are rapid, economical and reliable genotyping tools. Non‐heading Chinese cabbage (Brassica rapa L. subsp. chinensis Makino) is now an economically important vegetable crop worldwide. In this study, 1,167 SNPs were evaluated for 7polymorphism among 70 representative non‐heading Chinese cabbage inbred lines using a Kompetitive Allele Specific PCR (KASP) genotyping assay. On the basis of identified polymorphisms and the results of a principal component analysis, we selected 50 core SNPs that were balanced sufficiently to provide adequate information for genetic identification. The core SNPs were used for construction of a neighbour‐joining dendrogram that separated the 70 inbred lines into four main groups and several subgroups corresponding to Caixin, Heiyebaicai, Huangxinwu, Naibaicai, Taitsai, Pak‐choi, and Wutatsai. This categorization was superior to that achieved using a dataset of 479 polymorphic SNPs. To confirm the utility of the core SNP markers in genetic identification, we tested their stability and resolution using 162 commercial hybrid cultivars. The SNPs, which represent a cost‐effective, accurate marker set for germplasm analysis and cultivar identification, are suitable for molecular marker‐assisted breeding in non‐heading Chinese cabbage.     

Revisiting rice breeding methods – evaluating the use of rapid generation advance (RGA) for routine rice breeding

Rice production needs to increase in the future in order to meet increasing demands. The development of new improved and higher yielding varieties more quickly will be needed to meet this demand. However, most rice breeding programmes in the world have not changed in several decades. In this article, we revisit the evidence in favour of using rapid generation advance (RGA) as a routine breeding method. We describe preliminary activities at the International Rice Research Institute (IRRI) to re-establish RGA on a large scale as the main breeding method for irrigated rice breeding. We also describe experiences from the early adoption at the Bangladesh Rice Research Institute. Evaluation of RGA breeding lines at IRRI for yield, flowering time and plant height indicated transgressive segregation for all traits. Some RGA lines were also higher yielding than the check varieties. The cost advantages of using RGA compared to the pedigree method were also empirically determined by performing an economic analysis. This indicated that RGA is several times more cost effective and advantages will be realized after 1 year even if facilities need to be built. Based on our experience, and previous independent research empirically testing the RGA method in rice, we recommend that this method should be implemented for routine rice breeding in order to improve breeding efficiency.     

大豆对大豆胞囊线虫侵染的应答机制研究

大豆胞囊线虫病(Heterodera glycines,soybean cyst nematode,SCN)是大豆生产上的重要病害,其特点为危害重、分布广、难防治,每年对大豆生产造成极大的损失。种植大豆抗性新品种是防治SCN目前最为有效的措施,研究大豆对SCN侵染的应答机制,是培育大豆持久抗病品种的前提,对加快抗线虫品种选育及SCN的防控具有重要的意义。本文综述了大豆对SCN侵染的组织细胞学应答机制;介绍了大豆在SCN侵染后酶系变化及酚类代谢的生理生化应答机制;从分子水平阐明了SCN侵染后大豆的基因转录变化,差异蛋白及DNA甲基化的应答机制,以期为大豆胞囊线虫病害的进一步研究与防治提供参考。     

Identification,characterization and full-length sequence analysis of a novel endornavirus in common sunflower (Helianthus annuus L.)

To identify the possible quarantine viruses in seven common sunflower varieties imported from the United States of America and the Netherlands, we tested total RNAs extracted from the leaf tissues using next-generation sequencing of small RNAs. After analysis of small RNA sequencing data, no any quarantine virus was found, but a double-stranded RNA(dsRNA) molecule showing typical genomic features of endornavirus was detected in two varieties, X3939 and SH1108. Full-length sequence and phylogenetic analysis showed that it is a novel endornavirus, temporarily named as Helianthus annuus alphaendornavirus(HaEV). Its full genome corresponds to a 14 662-bp dsRNA segment, including a 21-nt 5′ untranslated region(UTR), 3' UTR ending with the unique sequence CCCCCCCC and lacking a poly(A) tail. An open reading frame(ORF) that encodes a deduced 4 867 amino acids(aa) polyprotein with three domains: RdRP, Hel and UGT(UDP-glycosyltransferase). HaEV mainly distributed in the cytoplasm but less in the nucleus of leaf cells by fluorescence in situ hybridization(FISH) experiment. This virus has a high seed infection rate in the five varieties, X3907, X3939, A231, SH1108 and SR1320. To our knowledge, this is the first report about the virus of the family Endornaviridae in the common sunflower.     

Establishing an eyeball-weight relationship for Litopenaeus vannamei using machine vision technology

This study aimed to establish a shrimp eyeball-weight relationship model for Litopenaeus vannamei using machine vision technology. A total of 295 shrimp were sampled from a recirculating aquaculture system (RAS). The long-axis length (d), body length (L), and body weight (W) of each individual was measured. The long axis length of the shrimp eyeball was identified and measured using machine vision technology. Continuous fitting and piecewise fitting models were used to construct the eyeball-weight relationship model for L. vannamei. The continuous fitting relationship model was described as: W = 38.865d^2.7914, while the piecewise model was described as: d < 2 mm, W = 0.0326d^3.7363, R² = 0.9288; 2 mm ≤ d < 3.9 mm, W = 0.0401d^3.104, R² = 0.9629; 3.9 mm ≤ d < 5.8 mm, W = 0.0421d^3.0311, R² = 0.9216; 5.8 mm < d, W = 0.103d^2.6226, R² = 0.9457. The root mean square error (RMSE) of the piecewise fitting model (0.0244, 0.1575, 0.5034, 0.7072) was smaller than the continuous fitting model (0.8229). The correlation coefficient (R²) of the piecewise model (0.9288, 0.9629, 0.9216, and 0.9457) was similar to that of the continuous fitting model (R² = 0.9621). The results indicated that the piecewise fitting model is suitable for calculating the biomass of L. vannamei in RAS and provides a novel way of estimating the biomass of L. vannamei cultured in RAS. The piecewise fitting model can also provide the foundation of evaluating the production of shrimp using underwater image recognition in intelligent aquaculture systems.     

东北三省大豆种质品质性状鉴评与综合分析

根据目标性状有的放矢的选配杂交亲本是提高优异品质组成品种的选择效率的基本前提。本研究对东北三省102份大豆种质资源的蛋白、氨基酸组分、油份及脂肪酸组分进行测定,通过遗传多样性、主成分和聚类分析,对其进行表型鉴定及基因型分类以综合评价种质品质特性。结果表明:东北三省大豆种质油份及脂肪酸组分变异较丰富,遗传多样性程度较高。根据主成分分析筛选到9个主成分进行聚类分析,通过聚类分析将供试种质资源分为5类。第I类群蛋白含量较高、油份含量偏低,第II类群蛋白、油份含量均居中,第III类群油份含量较高、蛋白含量偏低,第IV类群高油,第V类群高蛋白,类群间的氨基酸、脂肪酸组分各有差异。需根据育种目标在群体间选配亲本,以提高品质育种的效率。     

一种基于RPA的番茄褪绿病毒检测方法

番茄褪绿病毒Tomato chlorosis virus(ToCV)引起番茄褪绿病毒病,给番茄生产造成严重危害。开发快速准确的检测方法对该病害的防控具有重要意义。利用番茄褪绿病毒外壳蛋白(CP)基因序列,设计特异性引物,建立了ToCV的重组酶聚合酶等温扩增(recombinase polymerase amplification, RPA)检测方法,同时分析了该方法的灵敏度和特异性。结果表明,建立的ToCV-RPA方法在38℃恒温下40 min可从ToCV阳性的番茄样品中扩增出246 bp的特异性条带。扩增时间短,对设备要求低,且与番茄其他病毒无交叉反应,特异性好,灵敏度可达到PCR方法的10倍,适用于ToCV的快速检测。