分子标记辅助选择改良水稻恢复系香5的病虫抗性

香5是由湖北省农科院选育的优质两系杂交稻恢复系,所配组合广两优5号(广占63-4S/香5)于2013年通过了湖北省审定.利用回交和分子标记辅助选择技术,将供体亲本MD12086-1351中的抗稻瘟病基因Pi9、抗褐飞虱基因Bph14、Bph15和抗白叶枯病基因Xa23渗入到香5背景中,育成了3个同时携带Pi9、Bph14、Bph15和Xa23基因的新株系.鉴定结果表明,新株系的叶瘟抗性明显提高,穗颈瘟抗性部分提高,苗期抗褐飞虱,分蘖盛期高抗白叶枯病;产量、主要农艺性状、香味和稻米品质主要指标与香5相似.新株系所配的组合在产量、主要农艺性状上与香5所配的组合相似.表明新株系可以作为香5的替代系用于培育抗稻瘟病、抗褐飞虱和抗白叶枯病的两系杂交稻新组合.     

Comparative proteomics analysis of maize (Zea mays) leaves infected by small brown planthopper (Laodelphax striatellus)

Maize rough dwarf disease (MRDD) is a viral disease caused by brown planthopper infestation, and leads to great yield loss, especially in China. Comparative proteomics was performed using maize inbred line Zheng 58 and LN 287. MRDD pathogen was detected as rice black-streaked dwarf virus (RBSDV) by quantitative real time PCR (qRT-PCR) in Shandong Province, China. The modified trichloroacetic acid (TCA)/acetone method was used for soluble protein extraction from leaves. Two-dimensional electrophoresis (2-DE) analysis was performed on 24-cm long, pH 4–7 linear immobilized pH gradient (IPG) strips, and gels were stained with silver and coomassie brilliant blue. We identified 944 proteins expressed in RBSDV infected maize leaves by proteomics approaches. Among these, 44 protein spots that revealed a 1.5-fold difference in intensity were identified by mass spectrometry between mock-inoculated and RBSDV infected samples. Among these, 17 and 26 spots were up-regulated, and 27 and 18 spots were down-regulated in the virus infected samples of Zheng 58 and LN 287, respectively. Differential protein spots were analyzed by mass spectrometry identification, which could be divided into six categories. Furthermore, the expression of stress-related proteins was detected and confirmed by qRT-PCR. This study lays the foundation for further investigations, enabling the enhancement of MRDD resistance in maize.     

湘桂走廊白背飞虱早期虫源及灾变大气背景

为阐明湘桂走廊白背飞虱早期虫源及大气背景在其北迁过程中的影响,利用HYSPLIT软件模拟白背飞虱灾变性迁入高峰期、始见期后向轨迹,分别从个性及共性角度,分析湘桂走廊白背飞虱主要虫源地及其灾变大气背景。结果表明：（1）1991—2014年湘桂走廊白背飞虱的迁入始见期、首次迁入高峰期分别为0.9 d/10 a、1.6 d/10 a;（2）西南路径是湘桂走廊白背飞虱初始虫源迁入的主要路径,永福站、兴安站和全州站虫源西南路径占比分别为54%、50%、72%;（3）中南半岛白背飞虱借助微弱上升气流完成起飞,850 hPa高度上的强劲的西南气流、丰富的水汽条件有利于其北迁,降水是白背飞虱降落的重要大气动力机制。研究结果为白背飞虱的异地监测及科学防治提供科学依据。     

两个褐飞虱海藻糖转运蛋白基因的结构及调控海藻糖代谢功能

【目的】海藻糖是昆虫中的主要血糖物质,在昆虫的发育及生理活动中发挥重要功能。其中,海藻糖转运蛋白（Tret）在将海藻糖从其生成组织（例如脂肪体）运输到其消耗组织的过程中起着重要作用。本研究通过分析褐飞虱（Nilaparvata lugens）两条Tret1的序列结构,进一步抑制NlTret1的表达,探讨这两个NlTret1在褐飞虱体内的生物学功能。【方法】以褐飞虱两条Tret1序列为研究对象,利用生物信息学技术分析其蛋白结构以及与其他昆虫之间的同源性。采用RNAi（RNA interference）技术将合成的外源dsRNA（double-stranded RNA）注射到实验室饲养褐飞虱种群体内,抑制其体内NlTret1的表达,分别在注射48 h后取材,抽取总RNA并用反转录试剂盒合成第一链cDNA,采用qRT-PCR（quantitative real-time PCR）技术检测dsNlTret1的干扰效果以及RNAi后褐飞虱体内海藻糖代谢通路中相关基因的表达,最后测定葡萄糖、海藻糖和糖原含量以及海藻糖酶活性。【结果】生物信息学分析表明,NlTret1-like X1和NlTret1-2 X1的开放阅读框长度分别为1 920和1 578 bp,分别编码639和525个氨基酸,预测蛋白分子量分别为69.29和58.71 kD,等电点分别为8.32和8.36;NlTret1-like X1和NlTret1-2 X1的二级结构主要包含螺旋和卷曲;保守结构域分析显示它们均属于MFS家族。进化分析结果显示,不同昆虫的Tret1蛋白具有较高的同源性,且褐飞虱与其他半翅目昆虫具有较近的亲缘关系;与注射dsGFP组相比,注射靶标基因的dsRNA后均能够显著沉默本基因的表达;褐飞虱体内糖原、葡萄糖在注射dsNlTret1-like X1和dsNlTret1-2 X1后,其含量均无显著变化,然而不同于注射dsNlTret1-2 X1组,在注射dsNlTret1-like X1后褐飞虱体内海藻糖含量极显著升高;干扰NlTret1-like X1 48 h后褐飞虱体内TPS1、TPS2、TRE1-1、TRE1-2和TRE2的表达均极显著下调;而干扰NlTret1-2 X1 48 h后褐飞虱体内TPS1、TPS2和TRE1-1的表达虽也极显著下降,但TRE1-2和TRE2极显著上调;在注射dsNlTret1-like X1后,试虫可溶性海藻糖酶和膜结合型海藻糖酶的活性均极显著降低,而在注射dsNlTret1-2 X1后无显著变化。【结论】褐飞虱的两个Tret1在不同组织间发挥着不同的功能,其中NlTret1-like X1在特异性转运海藻糖参与能量供应中起到更为显著的作用。研究结果有助于探索Tret1在昆虫或无脊椎动物中调控海藻糖代谢平衡的调节机制,可为将来通过调控血糖平衡来控制褐飞虱等害虫提供理论依据。     

生物杀虫剂组合在有机水稻种植中的作用效果

针对上海市崇明区有机水稻生产过程中病虫害防治效果不佳的问题,在水稻无化肥、无化学农药种植条件下开展了生物杀虫剂对水稻主要害虫稻纵卷叶螟和稻飞虱的田间防控效果评价。在崇明区的2个试验基地,于水稻不同生长时期,以绿僵菌、白僵菌产品分别与病毒杀虫剂MbNPV、植物源杀虫剂苦参碱和细菌杀虫剂Bt组合进行施药处理。结果表明,应用绿僵菌/白僵菌与MbNPV组合,能够有效防治稻纵卷叶螟,在稻纵卷叶螟发生较早的基地,施药后7 d和14 d的防效可达58.2%～81.2%,而在稻纵卷叶螟发生较晚的另一基地,施药后7 d防效可达61.3%和65.6%;绿僵菌/白僵菌与苦参碱组合,可用于防治稻飞虱,7 d和14 d的防效达64.7%～76.1%;绿僵菌/白僵菌与苏云金杆菌组合防治稻纵卷叶螟的14 d防效分别达64.0%和75.4%,优于对稻飞虱的防效;各组合试验结果显示,白僵菌有相对较长的持效作用,且在害虫迁入早期阶段(水稻发育分蘖盛期至拔节期)施用生物农药比在中后期(抽穗期及之后)施用的防效高。可见,生物杀虫剂组合可作为农作物绿色防控的有效手段。     

卵磷脂桶混助剂的特性及其在防治稻飞虱中对氟啶虫胺腈的协同增效作用

为明确新型卵磷脂类桶混助剂“融透”的特性，采用透射电镜和激光粒度分析仪表征了其物理性质，并通过室内生物测定和田间药效试验测定了其在防治稻飞虱中对氟啶虫胺腈的协同增效作用。结果表明，“融透”中含有圆形脂质体，Z平均粒径为129.5 nm。室内生物测定结果表明，“融透”可使氟啶虫胺腈对褐飞虱的毒力增加1.67倍。田间药效试验表明，“融透”可使氟啶虫胺腈对稻飞虱的防效提高9.04%~41.77%；当氟啶虫胺腈减量40%时，添加“融透”的处理对稻飞虱的防效与未添加“融透”的常量组相当。研究结果表明，卵磷脂桶混助剂“融透”可以通过形成脂质体提高药剂的利用率，从而达到增效的作用。     

丁硫克百威对白背飞虱和南方水稻黑条矮缩病的防治研究

为明确丁硫克百威拌种剂对水稻白背飞虱和南方水稻黑条矮缩病的防治效果及其对天敌的影响,选择不同剂量的35%丁硫克百威种子处理干粉剂进行拌种。结果表明:剂量是30 g/kg种子拌种可有效防治白背飞虱,在播种后32 d、43 d、50d和57 d的防效分别为:90.6%、78.4%、67.9%和61.1%;同时通过防治白背飞虱减少传毒媒介,能有效控制南方水稻黑条矮缩病的危害,防效为76.9%。     

抗虫材料为主体的集成技术对稻飞虱的防控效果

本文研究了由新的抗虫材料‘中9A 07/WH1462’与控氮、晒田和天敌保护等措施有机结合的环境友好集成技术体系对稻飞虱的控制效果。结果表明,该技术体系在未施用化学农药的情况下,使稻飞虱等害虫种群数量得到有效控制,较好地保护了天敌,使稻田节肢动物群落结构得到一定优化。集成技术稻田的害虫所占比例比对照田降低了57.6%,其天敌所占比例比对照田增加了19.6%,同时集成技术稻田的水稻产量比对照田增加了10.9%。     

Development and Identification of Introgression Lines from Cross of Oryza sativa and Oryza minuta

Introgression line population is effectively used in mapping quantitative trait loci(QTLs),identifying favorable genes,discovering hidden genetic variation,evaluating the action or interaction of QTLs in multiple conditions and providing the favorable experimental materials for plant breeding and genetic research.In this study,an advanced backcross and consecutive selfing strategy was used to develop introgression lines(ILs),which derived from an accession of Oryza minuta(accession No.101133) with BBCC genome,as the donor,and an elite indica cultivar IR24(O.sativa),as the recipient.Introgression segments from O.minuta were screened using 164 polymorphic simple sequence repeat(SSR) markers in the genome of each IL.Introgressed segments carried by 131 ILs covered the whole O.sativa genome.The average number of homozygous O.minuta segments per introgression line was about 9.99.The average length of introgressed segments was approximate 14.78 cM,and about 79.64% of these segments had sizes less than 20 cM.In the genome of each introgression line,the O.minuta chromosomal segments harbored chromosomal fragments of O.sativa ranging from 1.15% to 27.6%,with an overall average of 8.57%.At each locus,the ratio of substitution of O.minuta alleles had a range of 1.5% 25.2%,with an average of 8.3%.Based on the evaluation of the phenotype of these ILs,a wide range of alterations in morphological and yield-related traits were found.After inoculation,ILs 41,11 and 7 showed high resistance to bacterial blight,brown planthopper and whitebacked planthopper,respectively.These O.minuta-O.sativa ILs will serve as genetic materials for identifying and using favorable genes from O.minuta.