云南地方稻种持久抗瘟性的研究

1996～1999年,在不同纬度、不同海拔和不同稻作生态类型的重病区设立5个持久抗性稻瘟病鉴定圃。试验材料为云南的74份地方稻种资源,其中粳稻56份,籼稻18份(含野生稻3份)。通过多个抗性组分进行了系统研究,初步表明大白谷(粳、墨江县)、毫弄早(籼、勐海县)、毫玉浪(籼、勐海县)、疣粒野生稻(野、西双版纳自治州)等具有持久抗瘟性能;其中疣粒野生稻高抗细菌性条斑病,对白叶枯病抗性为0级,接近免疫,中抗稻瘟病。通过对品种多抗性组分分析和品种抗性系统聚类分析,提出在不同生态类型时、空动态的病叶片上的产孢量和病斑表型可作为简易、快速鉴定持久抗瘟性指标。     

云南疣粒野生稻稻瘟病抗性

野生稻(Oryza rufipogo)保存有许多栽培稻(O. sativa)不具备或已经消失的优异基因资源, 是扩大栽培稻遗传背景、改良产量与品质、提高抗病虫害及抗逆境能力的重要基因库。疣粒野生稻(O. meyeriana)是中国3种野生稻资源之一, 主要分布在云南。为进一步了解其稻瘟病抗性, 首先利用来自不同稻作区的稻瘟病菌株, 通过注射接种法对疣粒野生稻进行系统的稻瘟病抗性鉴定, 发现疣粒野生稻对接种的所有稻瘟病菌株都感病。进一步采用3'/5' RACE方法, 从疣粒野生稻中克隆了水稻同源基因Pid2和Pid3, 并构建过表达转基因株系对基因功能进行了研究。结果表明, Pid2和Pid3与疣粒野生稻中同源基因间在DNA和氨基酸水平上有较大的序列差异, 过表达转基因的日本晴植株对稻瘟病菌的敏感性与对照相似。推测疣粒野生稻在自然接种条件下, 表现出的抗稻瘟病表型很可能是其旱生叶片结构特征形成了对稻瘟病菌侵染的天然屏障。对控制疣粒野生稻这一类性状基因资源的挖掘和利用, 有利于优良抗性水稻品种的培育。研究结果为疣粒野生稻的研究利用提供了新信息和新思路。     

海南普通野生稻稻瘟病抗性资源调查和鉴定

为挖掘海南普通野生稻稻瘟病抗性资源,2010年诱发鉴定了41个居群410份材料苗期叶瘟,2011年接种稻瘟病菌（YC25）鉴定了37个居群121份材料穗颈瘟,2012年调查了80个居群2461份材料田间自然状态的抗病性。结果表明：苗期叶瘟抗性鉴定有21份高抗、117份抗病。苗期抗、高抗叶瘟性的138份材料中穗颈瘟鉴定4份高抗和3份抗病,14份表现为田间自然抗病。苗期叶瘟鉴定不抗病或未作此鉴定材料中,4份表现为抗穗颈瘟和田间自然抗病。这些抗性材料来自海口、文昌、万宁、三亚、澄迈、东方等地。本研究为海南普通野生稻资源进一步研究和抗稻瘟病育种利用提供参考     

海南普通野生稻稻瘟病的抗性鉴定与评价

在苗期应用自然诱发鉴定法对海南普通野生稻(Oryza rufipogonGriff.)41个居群的410份材料进行了2年的稻瘟病(rice blast)抗性鉴定,结果表明:经过初鉴和复鉴,410份海南普通野生稻中有21份表现高抗,占5.1%,117份表现抗,占28.5%,说明海南普通野生稻具有较好的稻瘟病抗性。     

抗稻瘟病水稻资源抗性基因Pita、Pib、Pi9以及Pikm的分布研究

利用抗稻瘟病水稻资源品种杂交,聚合多个抗性基因是培育持久抗稻瘟病水稻新品种的主要育种途径.利用分子标记技术对水稻抗性资源进行基因型鉴定是分子辅助聚合育种的基础.通过以亚华种业科学院稻瘟病病圃抗病水稻资源为材料,利用特异性分子标记对Pi9、Pita、Pib以及Pikm基因在水稻抗稻瘟病资源的分布进行了鉴定,初步建立了抗性基因数据库.同时对抗性基因及与抗性反应的相关性进行了探讨,结果表明以Pi9为主效基因,同时聚合Pita和Pib抗性基因能提高持久抗稻瘟病能力.     

小麦品种抗麦红吸浆虫鉴定与抗性分析

利用一套较完整的田间自然感虫鉴定法,对183份小麦品种进行了抗麦红吸浆虫鉴定.鉴定结果表明,小麦不同品种对麦红吸浆虫的抗性存在着显著差异:高抗、中抗、低抗、感虫、高感的小麦品种,分别占参试品种的24.59%、16.94%、18.58%、14.21%、25.68%.不同抗性类型的小麦品种其穗被害率、粒被害率、估计损失率、抗性指数均有较大的差异,其中表现高抗类型45份,中抗类型31份.这些品种既可作为麦红吸浆虫发生区控制麦红吸浆虫危害的主推品种和后备品种,也可作为亲本材料提供给育种单位利用.     

稻瘟病抗性基因Pita和Pib在我国水稻主栽品种中的分布

主效抗稻瘟病基因Pita和Pib在我国很多稻区表现高水平的稻瘟病抗性,被广泛应用于我国的水稻育种和生产.但这2个基因在国内主栽品种中的分布及利用情况一直缺乏详细的资料,致使育种利用上存在着盲目性.本研究利用源于Pita和Pib基因本身的特异性分子标记,结合稻瘟病菌接种鉴定,检测和分析了我国58份水稻主栽品种(杂交稻亲本)的Pita和Pib抗性基因型.结果表明,特籼占25、佳禾早占、密阳46、测64-7等4个籼稻品种携带Pita和Pib 2个基因;籼小占等4个籼稻品种(系)和早丰9号等5个粳稻品种携带抗性基因Pita;绵恢501等5个籼稻品种(系)和粳稻品种武育粳7号、辽粳454携带抗性基因Pib.     

野生稻细菌性条斑病抗性资源筛选及遗传分析

对1655份普通野生稻(Oryza rufipogon Griff.)和31份药用野生稻(O. officinalis Wall. ex Watt)进行了细菌性条斑病(Xanthomonas oryzae pv. oryzicola,简称细条病)抗性鉴定,结果发现在普通野生稻中有57份抗病材料,其中3级抗性有31份,占总数的1.87%;5级中抗有26份,占总数1.57%.在药用野生稻资源中,有15份抗病材料,占总数的48.4%.选取了8份普通野生稻抗性资源(分别命名为DP1、DP3、DP5、DP9、DP15、DP16、DP17和DP20)与9311杂交,再自交或与9311回交后,分别获得BC1、F1和F2后代.在接种鉴定中发现这些抗性资源的BC1或F1所有植株均对细条病表现感病,说明这8份材料的抗性属于隐性遗传.在DP3与9311杂交的F2群体中,抗感植株的分离比符合1∶ 15的比例,说明DP3的抗性由2对隐性重叠作用基因控制.研究结果表明,在野生稻中可以获得一批具有较大利用价值的细条病抗性资源,其中药用野生稻资源中抗性材料所占的比例较大.     

我国蚕豆品种资源对蚕豆锈病的抗性鉴定

1998～2000年对241份蚕豆品种(系)进行抗蚕豆锈病鉴定,其中表现高抗(HR)的材料2份,占0.83%;抗病(R)和中抗(MR)的材料131份,占54.36%.中感(MS)和感病(S)材料104份,占43.15%,高感(HS)的材料4份,占1.66%.结果表明,不同地理来源、株高、单株粒数、单株产量及粒重的材料抗性有差异.     

甘薯兼抗薯瘟病和蔓割病种质筛选鉴定

用甘薯瘟2个致病型菌株对557份甘薯品种(系)分菌系做盆苗和田间接种鉴定,并用蔓割病菌接种鉴定.结果表明,筛选出兼抗薯瘟病2个菌系的品种(系)31份,兼抗甘薯瘟和蔓割病的品种(系)有16份,其中泉薯860、泉薯854、金山93、榕选416、金山908、广薯88-70和泉薯853等7份与我省主栽抗病品种湘薯75-55(CK)相比,具有类似的双抗性能力,而其抗瘟能力更强.福薯87、林泗2号、金山57和湛93-16等4份对照品种只抗单个病害或兼抗单个菌系.     

Expression of a harpin-encoding gene in rice confers durable nonspecific resistance to Magnaporthe grisea

Engineering durable nonspecific resistance to phytopathogens is one of the ultimate goals of plant breeding. However, most attempts to reach this goal fail as a result of rapid changes in pathogen populations and the sheer diversity of pathogen infection mechanisms. In this study, we show that the expression of a harpin-encoding gene ( hrf1 ), derived from Xanthomonas oryzae pv. oryzae, confers nonspecific resistance in rice to the blast fungus Magnaporthe grisea . Transgenic plants and their T₁–T₇ progenies were highly resistant to all major M. grisea races in rice-growing areas along the Yangtze River, China. The expression of defence-related genes was activated in resistant transgenic plants, and the formation of melanized appressoria, which is essential for foliar infection, was inhibited on plant leaves. These results suggest that harpins may offer new opportunities for generating broad-spectrum disease resistance in other crops.     

转拟南芥AtNPR1基因增强水稻对水稻白叶枯病和稻瘟病的抗性

AtNPR1基因是拟南芥系统获得抗性的一个重要调节基因,在拟南芥中过量表达AtNPR1基因能使拟南芥对细菌和真菌的抗性同时增强.为了研究在水稻中过量表达AtNPR1基因对水稻抗病性的影响,将该基因转入到广西主栽籼稻恢复系品种桂99中.经PCR验证得到了79株转基因植株,DNA斑点杂交表明ATNPR1基因已经整合到桂99染色体DNA中.Northern杂交和RT-PCR分析表明,AtNPR1基因在桂99中已经表达;同时还检测了转基因植株对水稻白叶枯病和稻瘟病的抗性,结果表明转基因植株对该两种病害的抗性均显著增强.     

Screening for resistance against Pseudomonas syringae in rice-FOX Arabidopsis lines identified a putative receptor-like cytoplasmic kinase gene that confers resistance to major bacterial and fungal pathogens in Arabidopsis and rice

Approximately 20,000 of the rice-FOX Arabidopsis transgenic lines, which overexpress 13,000 rice full-length cDNAs at random in Arabidopsis, were screened for bacterial disease resistance by dip inoculation with Pseudomonas syringae pv. tomato DC3000 (Pst DC3000). The identities of the overexpressed genes were determined in 72 lines that showed consistent resistance after three independent screens. Pst DC3000 resistance was verified for 19 genes by characterizing other independent Arabidopsis lines for the same genes in the original rice-FOX hunting population or obtained by reintroducing the genes into ecotype Columbia by floral dip transformation. Thirteen lines of these 72 selections were also resistant to the fungal pathogen Colletotrichum higginsianum. Eight genes that conferred resistance to Pst DC3000 in Arabidopsis have been introduced into rice for overexpression, and transformants were evaluated for resistance to the rice bacterial pathogen, Xanthomonas oryzae pv. oryzae. One of the transgenic rice lines was highly resistant to Xanthomonas oryzae pv. oryzae. Interestingly, this line also showed remarkably high resistance to Magnaporthe grisea, the fungal pathogen causing rice blast, which is the most devastating rice disease in many countries. The causal rice gene, encoding a putative receptor-like cytoplasmic kinase, was therefore designated as BROAD-SPECTRUM RESISTANCE 1. Our results demonstrate the utility of the rice-FOX Arabidopsis lines as a tool for the identification of genes involved in plant defence and suggest the presence of a defence mechanism common between monocots and dicots.     

Molecular evolution of the rice blast resistance gene Pi-ta in invasive weedy rice in the USA

The Pi-ta gene in rice has been effectively used to control rice blast disease caused by Magnaporthe oryzae worldwide. Despite a number of studies that reported the Pi-ta gene in domesticated rice and wild species, little is known about how the Pi-ta gene has evolved in US weedy rice, a major weed of rice. To investigate the genome organization of the Pi-ta gene in weedy rice and its relationship to gene flow between cultivated and weedy rice in the US, we analyzed nucleotide sequence variation at the Pi-ta gene and its surrounding 2 Mb region in 156 weedy, domesticated and wild rice relatives. We found that the region at and around the Pi-ta gene shows very low genetic diversity in US weedy rice. The patterns of molecular diversity in weeds are more similar to cultivated rice (indica and aus), which have never been cultivated in the US, rather than the wild rice species, Oryza rufipogon. In addition, the resistant Pi-ta allele (Pi-ta) found in the majority of US weedy rice belongs to the weedy group strawhull awnless (SH), suggesting a single source of origin for Pi-ta. Weeds with Pi-ta were resistant to two M. oryzae races, IC17 and IB49, except for three accessions, suggesting that component(s) required for the Pi-ta mediated resistance may be missing in these accessions. Signatures of flanking sequences of the Pi-ta gene and SSR markers on chromosome 12 suggest that the susceptible pi-ta allele (pi-ta), not Pi-ta, has been introgressed from cultivated to weedy rice by out-crossing.     

中国普通野生稻遗传分化的RAPD研究

多数学者已认定亚洲栽培稻（ＯｒｙｚａｓａｔｉｖａＬ．）的祖先是普通野生稻（Ｏ．ｒｕｆｉｐｏｇｏｎ）。然而栽培稻的籼、粳分化是发生在驯化之前还是在驯化之后,也即普通野生稻是否存在籼、粳分化的问题,是十几年来稻作起源研究中争论的热点之一。Ｓｅｃｏｎｄ［１］用多个同工酶位点的分析结果得出结论,普通野生稻在驯化为栽培稻之前就已经发生了籼、粳分化,即有籼型普通野生稻和粳型普通野生稻之分。Ｍｏｒｉｓｈｉｍａ和Ｇａｄｒｉｎａｂ［２］用２４个形态和生理性状及１２个同工酶位点和杂交亲合力等方法证明普通野生稻没有发…     

Brassinosteroid functions in a broad range of disease resistance in tobacco and rice

Brassinolide (BL), considered to be the most important brassinosteroid (BR) and playing pivotal roles in the hormonal regulation of plant growth and development, was found to induce disease resistance in plants. To study the potentialities of BL activity on stress responding systems, we analyzed its ability to induce disease resistance in tobacco and rice plants. Wild-type tobacco treated with BL exhibited enhanced resistance to the viral pathogen tobacco mosaic virus (TMV), the bacterial pathogen Pseudomonas syringae pv. tabaci (Pst), and the fungal pathogen Oidium sp. The measurement of salicylic acid (SA) in wild-type plants treated with BL and the pathogen infection assays using NahG transgenic plants indicate that BL-induced resistance does not require SA biosynthesis. BL treatment did not induce either acidic or basic pathogenesis-related (PR) gene expression, suggesting that BL-induced resistance is distinct from systemic acquired resistance (SAR) and wound-inducible disease resistance. Analysis using brassinazole 2001, a specific inhibitor for BR biosynthesis, and the measurement of BRs in TMV-infected tobacco leaves indicate that steroid hormone-mediated disease resistance (BDR) plays part in defense response in tobacco. Simultaneous activation of SAR and BDR by SAR inducers and BL, respectively, exhibited additive protective effects against TMV and Pst, indicating that there is no cross-talk between SAR- and BDR-signaling pathway downstream of BL. In addition to the enhanced resistance to a broad range of diseases in tobacco, BL induced resistance in rice to rice blast and bacterial blight diseases caused by Magnaporthe grisea and Xanthomonas oryzae pv. oryzae, respectively. Our data suggest that BDR functions in the innate immunity system of higher plants including dicotyledonous and monocotyledonous species.     

Manipulating broad-spectrum disease resistance by suppressing pathogen-induced auxin accumulation in rice

Breeding crops with the quality of broad-spectrum disease resistance using genetic resources is one of the principal goals of crop improvement. However, the molecular mechanism of broad-spectrum resistance remains largely unknown. Here, we show that GH3-2, encoding an indole-3-acetic acid (IAA)-amido synthetase, mediates a broad-spectrum resistance to bacterial Xanthomonas oryzae pv oryzae and Xanthomonas oryzae pv oryzicola and fungal Magnaporthe grisea in rice (Oryza sativa). IAA, the major form of auxin in rice, results in rice more vulnerable to the invasion of different types of pathogens, which is at least partly due to IAA-induced loosening of the cell wall, the natural protective barrier of plant cells to invaders. X. oryzae pv oryzae, X. oryzae pv oryzicola, and M. grisea secrete IAA, which, in turn, may induce rice to synthesize its own IAA at the infection site. IAA induces the production of expansins, the cell wall-loosening proteins, and makes rice vulnerable to pathogens. GH3-2 is likely contributing to a minor quantitative trait locus for broad-spectrum resistance. Activation of GH3-2 inactivates IAA by catalyzing the formation of an IAA-amino acid conjugate, which results in the suppression of expansin genes. Thus, GH3-2 mediates basal resistance by suppressing pathogen-induced IAA accumulation. It is expected that, regulated by a pathogen-induced strong promoter, GH3-2 alone may be used for breeding rice with a broad-spectrum disease resistance.     

RAPD Determinations on the Genetic Differentiation of Chinese Common Wild Rice

Ninety accessions which included Chinese common wild rice (Oryza rufipogon) from 8 provinces and traditional cultivars from lower and middle basins of Yangtze River, southeast of China and Yunnan Province as well as some commercial varieties were analyzed by RAPD with 24 primers. A scattered figure suggesting the indica-japonica and wild-domestication differentiations among 90 rice accessions was generated based on RAPD data. The results indicated that Chinese common wild rice, indica and japonica accessions were divided into 3 groups respectively. Chinese common wild rice were somewhat closer to the japonica type than the indica type.     

水稻长日抑制基因PhyB的多样性及区域适应性初探

Phy 是在长日照条件下抑制水稻开花的关键基因,但目前对水稻PhyB基因的遗传基础还不清楚,研究其分子遗传机制,对于培育光周期适应性广的品种以及扩大水稻种植区域具有重要意义。本研究选择78份亚洲栽培稻(34份籼稻和44份粳稻)及47份野生稻进行测序,对Phy B基因的核苷酸多态性、单倍型进行分析,计算籼稻、粳稻和野生稻的遗传多样性。结果表明,Phy B基因共有28个单倍型,其中有2个高频率的单倍型分别存在于2个栽培稻亚种中。从Network图可以看出栽培稻分为2组(A组和B组),A组栽培稻包括全部的籼稻和4个粳稻个体,B组栽培稻全是粳稻品种。亲缘地理学分析发现,A、B两组栽培稻具有明显不同的地理分布格局,且A组和B组开花时间差异显著,说明Phy B基因的2个高频率单倍型在2个栽培稻亚种中具有区域适应性,Phy B基因在栽培稻中具有明显的驯化信号,随着水稻种植区域的扩大,进化出适应不同地域特有的等位基因,导致开花时间对不同地区的区域适应性及多样性。