聚合白背飞虱和褐飞虱抗性基因创制杂交水稻恢复系

【目的】为了创制兼抗白背飞虱和褐飞虱的水稻恢复系,【方法】分别以抗褐飞虱材料B5(携带褐飞虱抗性基因Bph14和Bph15)及携带白背飞虱抗性位点qsI-4的籼型恢复系福恢7011为供体亲本,以骨干恢复系福恢676为轮回亲本,应用低世代分离群体田间表型结合单株鉴定与高世代稳定株系室内筛选和分子标记辅助选择相结合的方法,并对抗虫株系及其测交后代进行考查和农艺性状分析。【结果】选育出聚合Bph14、Bph15和qsI-4的恢复系材料3份,携带2个抗虫基因的恢复系材料3份。其中6份恢复系的褐飞虱抗性鉴定结果均表现中抗以上。通过抗性鉴定和杂交后代农艺性状分析筛选出具有生产应用潜力的恢复系材料2份。【结论】为褐飞虱和白背飞虱抗性聚合新种质的创制和应用提供了基础材料。     

应用DNA标记分析稻飞虱的抗性基因

简要地回顾了水稻抗飞虱的遗传位点定位和作图的新进展.来自具有不同基因组的野生稻渗入系的4个抗褐飞虱基因Bph 1、 bph 2、 bph 4和Bph 9,以及4个暂定名抗褐飞虱基因Bph 10(t)、bph 11(t)、bph 12(t)和Bph 13(t),目前已被定位于水稻12条染色体中的5条.其中,Bph 1、 bph 2、 Bph 9和Bph 10(t)在水稻第12染色体的长臂上形成1个连锁区段,位于bph 2位点附近约25 cM.检测出几个对田间抗性和杀卵作用有影响的QTL.抗白背飞虱基因Wbph 1、 Wbph 2和Wbph 6(t)已经或暂时定位了.粳稻中对白背飞虱具有杀卵抗性的QTL已进行了详细的分析,在第6染色体的短臂上检测到有效的QTL,在同一位点鉴定出1个显性的杀卵基因Ovc.在杀卵基因Ovc存在时,第1染色体上的1个QTL和第5染色体上的2个QTL增加白背飞虱的卵死亡率.用DNA标记进行QTL作图可以加深人们对作物抗虫性中复杂的生理和遗传机理的理解.标记辅助选择可以加速培育具多基因抗虫性的作物,还可以将野生种中的有利抗虫特性转入改良品种中,增加作物抗虫性的持久性和遗传多样性.     

聚合白背飞虱和褐飞虱抗性基因创制杂交水稻恢复系

【目的】为了创制兼抗白背飞虱和褐飞虱的水稻恢复系,【方法】分别以抗褐飞虱材料B5(携带褐飞虱抗性基因Bph14和Bph15)及携带白背飞虱抗性位点qsI-4的籼型恢复系福恢7011为供体亲本,以骨干恢复系福恢676为轮回亲本,应用低世代分离群体田间表型结合单株鉴定与高世代稳定株系室内筛选和分子标记辅助选择相结合的方法,并对抗虫株系及其测交后代进行考查和农艺性状分析。【结果】选育出聚合Bph14、Bph15和qsI-4的恢复系材料3份,携带2个抗虫基因的恢复系材料3份。其中6份恢复系的褐飞虱抗性鉴定结果均表现中抗以上。通过抗性鉴定和杂交后代农艺性状分析筛选出具有生产应用潜力的恢复系材料2份。【结论】为褐飞虱和白背飞虱抗性聚合新种质的创制和应用提供了基础材料。     

广谱抗稻瘟病种质75-1-127的褐飞虱抗性基因鉴定及分子标记辅助选择育种

【目的】水稻品系75-1-127携带广谱抗稻瘟病基因Pi9,已被广泛应用于抗稻瘟病水稻品种改良。笔者育种实践发现75-1-127表现出较强的褐飞虱抗性,因此鉴定该品系中的褐飞虱抗性基因并进行分子辅助选择育种。【方法】根据水稻品系B5中褐飞虱抗性基因Bph14和Bph15的序列,设计引物扩增75-1-127的基因组DNA,并对PCR产物进行测序分析。采用苗期集团法鉴定了75-1-127和B5的褐飞虱抗性表型。利用与Bph14与Bph15连锁的分子标记筛查了75-1-127为稻瘟病抗源回交转育的两系不育系后代,并鉴定了这些后代的稻瘟病抗性、褐飞虱抗性和主要农艺性状。【结果】75-1-127中含有与B5完全一致的Bph14和Bph15序列。75-1-127和B5苗期褐飞虱抗性均为1级。在以75-1-127为抗源改良的两系不育系中,携带Bph14、Bph15的单基因系或双基因系的褐飞虱抗性均得以改良,其中双基因聚合系的死苗率为8.5%,与供体亲本75-1-127以及阳性对照B5差异不显著,进一步证实75-1-127含有褐飞虱抗性基因。【结论】水稻品系75-1-127携带褐飞虱抗性基因Bph14和Bph15,可以作为抗源应用于水稻褐飞虱抗性育种。     

聚合白背飞虱和褐飞虱抗性基因创制杂交水稻恢复系

【目的】为了创制兼抗白背飞虱和褐飞虱的水稻恢复系，【方法】分别以抗褐飞虱材料B5（携带褐飞虱抗性基因Bph14和Bph15）及携带白背飞虱抗性位点qsI-4的籼型恢复系福恢7011为供体亲本，以骨干恢复系福恢676为轮回亲本，应用低世代分离群体田间表型结合单株鉴定与高世代稳定株系室内筛选和分子标记辅助选择相结合的方法，并对抗虫株系及其测交后代进行考查和农艺性状分析。【结果】选育出聚合Bph14、Bph15 和qsI-4的恢复系材料3份，携带2个抗虫基因的恢复系材料3份。其中6份恢复系的褐飞虱抗性鉴定结果均表现中抗以上。通过抗性鉴定和杂交后代农艺性状分析筛选出具有生产应用潜力的恢复系材料2份。【结论】为褐飞虱和白背飞虱抗性聚合新种质的创制和应用提供了基础材料。     

利用分子标记辅助选择改良水稻两系不育系C815S的褐飞虱抗性研究

以水稻品种B5(含褐飞虱抗性基因Bph14、Bph15)为供体亲本,目前生产上大面积应用的两系不育系C815S为受体亲本,利用分子标记辅助选择技术,将2个褐飞虱抗性基因Bph14和Bph15导入C815S,获得3个同时携带2个抗性基因的纯合改良株系。苗期群体鉴定和大田成熟期鉴定表明,3个改良不育株系的褐飞虱抗性均为3级,表现为抗;而受体亲本C815S为9级,表现为感。农艺性状考察表明,改良不育株系基本保持了受体亲本的优良性状,部分穗部性状还显著优于C815S。因此,利用分子标记辅助选择技术聚合Bph14和Bph15基因改良不育系褐飞虱抗性效果明显。     

水稻抗褐飞虱育种研究进展与展望

褐飞虱是危害最严重的水稻害虫之一,能降低产量并影响稻米品质。控制褐飞虱的关键在于利用品种自身的抗性基因培育新的抗虫品种。目前,已发现并报道了34个抗褐飞虱基因,其中28个主效基因已被定位,Bph3、Bph9、Bph14、Bph18、BPH18、Bph26、BPH29、Bph32和Bphi008a等基因已成功克隆。已有研究表明,聚合多个褐飞虱抗性基因的品种抗性明显高于含单个或不含抗性基因的品种,但目前抗性基因只有个别得到有效利用。本文对褐飞虱的生物型、抗性机制、抗性基因的定位和克隆以及在水稻育种上的应用进行了综述,并对抗褐飞虱育种面临的问题和育种对策进行了讨论。     

水稻抗褐飞虱基因研究进展与展望

褐飞虱严重影响水稻的产量和品质,抗虫品种的培育和种植是控制该虫害最安全有效的措施。水稻抗褐飞虱育种的关键是抗虫基因的发掘和合理利用。目前至少报道了34个褐飞虱抗性位点,其中28个主效抗性基因已被定位,显性基因Bph3、Bph14和Bph26已被成功克隆。介绍了水稻褐飞虱的生物型及抗性机理、褐飞虱抗性基因定位与克隆及抗虫基因在育种上的应用,并对水稻褐飞虱抗性育种面临的问题和应用前景进行了讨论。     

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.     

籼稻材料570011抗褐飞虱基因的遗传分析及鉴定

【目的】发掘籼稻570011中抗褐飞虱主效基因，为培育抗虫水稻新品种提供基因资源。【方法】采用苗期集团法对抗性亲本570011和感虫亲本9311杂交后代F3群体进行表型鉴定，结合F2群体基因型，使用作图软件构建染色体的局部遗传连锁图，对目标区段抗性位点进行检测和遗传效应评估。采用实时荧光定量PCR(qRT-PCR)分析定位区间内最可能的候选基因，并对其基因组序列进行测序，比较对应CDS和氨基酸序列并进行系统进化树分析。【结果】籼稻570011在苗期对褐飞虱表现高抗，且对褐飞虱有明显的抗生性和趋避性。统计发现F3群体抗虫株系数（抗性值<7）∶感虫株系数（抗性值≥7）为89∶35，卡方检验表明符合一对显性基因的分离规律。基因定位发现在第4染色体标记4M18.675和4M24.64之间的39 cM区域内检测到一个褐飞虱抗性位点，可能是已克隆基因BPH6的等位基因。qRT-PCR分析表明570011中Os04g35210(BPH6等位基因)在褐飞虱取食后表达量显著高于感虫材料9311。570011中Os04g35210基因与BPH6的CDS和氨基酸序列同源性分别达到99.08%和97.9...     

抗稻瘟病基因Pi9的STS连锁标记开发及在分子标记辅助育种中的应用

 利用带有广谱抗稻瘟病基因Pi9的籼稻品系75 1 127作为抗病基因供体亲本,用于扬稻6号和R6547抗病性的回交育种。通过比较75 1 127、扬稻6号和日本晴的Pi9基因位点的DNA序列,开发出了与Pi9基因紧密连锁的共显性STS（序列标记位点）标记PB9 1,用于Pi9基因的分子标记辅助选择。结合田间农艺性状选择和分子标记辅助选择,培育出8个Pi9基因纯合的回交后代株系。其中,具有扬稻6号和R6547遗传背景的株系各4个。经湖北恩施和宜昌的病圃鉴定,携带有抗病基因株系的稻瘟病抗性水平较受体品种扬稻6号和R6547有不同程度的提高。具有R6547遗传背景的株系08C893配制的杂交组合在上述病区的抗性表现也明显优于对照品种扬两优6号。上述结果说明,共显性标记PB9 1在Pi9抗稻瘟病基因分子标记辅助育种中具有应用价值,并且Pi9基因作为稻瘟病抗源之一可以在湖北稻区进行有效利用。     

RFLP and SSR mapping of a new gene for brown planthopper resistance introgressed from O. Eichingeri into cultivated rice(O. Sativa L.)

Nine brown planthopper(BPH) resistance genes have been registered so far, but of them only Bph1, bph2, Bph3, bph4, Bph9,and other three unregistered genes Bph10(t), Bph(t), bph(t) were located on chromosome 3, 4, 10, and 12, respectively, by using traditional and molecular mapping methods. To use the genes for BPH resistance in rice breeding and production, interspecific hybrids between cultivated rice and accessions of O. eichingeri (2n=24, CC), a wild rice species from Africa, with strong resistance to BPH and whitebacked planthopper were produced.     

Marker assisted pyramiding of <Emphasis Type="Italic">Bph6</Emphasis> and <Emphasis Type="Italic">Bph9</Emphasis> into elite restorer line 93–11 and development of functional marker for <Emphasis Type="Italic">Bph9</Emphasis>

Background

The brown planthopper (BPH) has become the most destructive and a serious threat to the rice production in Asia. Breeding the resistant varieties with improved host resistance is the most effective and ecosystem-friendly strategy of BPH biological management. As host resistance was always broken down by the presence of the upgrading BPH biotype, the more resistant varieties with novel resistance genes or pyramiding known identified BPH resistance genes would be needed urgently for higher resistant level and more durability of resistance.

Results

Here, we developed near isogenic lines of Bph9 (NIL-Bph9) by backcrossing elite cultivar 93–11 with Pokkali (harboring Bph9) using marker-assisted selection (MAS). Subsequently, we pyramided Bph6 and Bph9 in 93–11 genetic background through MAS. The resulting Bph6 and Bph9 pyramided line LuoYang69 had stronger antixenotic and antibiosis effects on BPH and exhibited significantly enhanced resistance to BPH than near isogenic lines NIL-Bph6 and NIL-Bph9. LuoYang69 derived hybrids, harboring heterozygous Bph6 and Bph9 genes, also conferred high level of resistance to BPH. Furthermore, LuoYang69 did not affect the elite agronomic traits and rice grain quality of 93–11. The current study also developed functional markers for Bph9. Using functional dominant marker, we screened and evaluated worldwide accessions of rice germplasm. Of the 673 varieties tested, 8 cultivars were identified to harbor functional Bph9 gene.

Conclusion

The development of Bph6 and Bph9 pyramided line LuoYang69 provides valuable resource to develop hybrid rice with highly and durable BPH resistance. The development of functional markers will promote MAS of Bph9. The identified Bph9 containing cultivars can be used as new sources for BPH resistance breeding programs.

     

Current Status of Brown Planthopper (BPH) Resistance and Genetics

Among the planthoppers of rice, the brown planthopper (BPH) is a major threat to rice production and causes significant yield loss annually. Host-plant resistance is an important strategy to reduce the damage caused by BPH and increase rice productivity. Twenty-one major genes for BPH resistance have been identified by using standard evaluation methods developed at the International Rice Research Institute (IRRI) to distinguish resistance or susceptibility of rice genotypes to BPH biotypes/populations. These genes are from diverse genetic resources such as land race cultivars and wild species of Oryza. Of the 21 resistance genes, 18 genes have been localized on specific region of six rice chromosomes using molecular genetic analysis and genomics tools. Some of these resistance genes are clustered together such as Bph1, bph2, Bph9, Bph10, Bph18, and Bph21 on the long arm of chromosome 12; Bph12, Bph15, Bph17 and Bph20 on the short arm of chromosome 4; bph11 and Bph14 on the long arm of chromosome 3 and Bph13(t) and bph19 on the short arm of chromosome 3. Six genes (Bph11, bph11, Bph12, bph12, Bph13 and Bph13) originated from wild Oryza species have either duplicate chromosome locations or wrong nomenclature. The discrepancy should be confirmed by allelism tests. Besides identification of major resistance genes, some quantitative trait loci (QTLs) associated with BPH resistance have also been identified on eight chromosomes. Most of the rice cultivars developed at IRRI possess one or two of the major resistance genes and the variety IR64 has many QTLs and confers strong resistance to BPH. More BPH resistance genes need to be identified from the wealth of gene pool available in the wild species of Oryza. Two BPH resistance genes (Bph14 and Bph18) have been cloned, and a snow drop lectin gene (GNA) has been identified and used in the development of BPH-resistant transgenic plants. Efficient introgression of resistance genes (Bph1, bph2, Bph3, Bph14, Bph15, Bph18, Bph20, and Bph21) into elite rice cultivars by marker-assisted selection together with strategic deployment of these genes can be an important approach to develop stable resistance to BPH and sustain rice production in the tropical and temperate rice growing regions.     

抗稻瘟病基因Pi9的STS连锁标记开发及在分子标记辅助育种中的应用

利用带有广谱抗稻瘟病基因Pi9的籼稻品系75-1-127作为抗病基因供体亲本,用于扬稻6号和R6547抗病性的回交育种。通过比较75-1-127、扬稻6号和日本晴的Pi9基因位点的DNA序列,开发出了与Pi9基因紧密连锁的共显性STS(序列标记位点)标记PB9-1,用于Pi9基因的分子标记辅助选择。结合田间农艺性状选择和分子标记辅助选择,培育出8个Pi9基因纯合的回交后代株系。其中,具有扬稻6号和R6547遗传背景的株系各4个。经湖北恩施和宜昌的病圃鉴定,携带有抗病基因株系的稻瘟病抗性水平较受体品种扬稻6号和R6547有不同程度的提高。具有R6547遗传背景的株系08C893配制的杂交组合在上述病区的抗性表现也明显优于对照品种扬两优6号。上述结果说明,共显性标记PB9-1在Pi9抗稻瘟病基因分子标记辅助育种中具有应用价值,并且Pi9基因作为稻瘟病抗源之一可以在湖北稻区进行有效利用。     

Understanding Brown Planthopper Resistance in Rice: Genetics,Biochemical and Molecular Breeding Approaches

Brown planthopper (BPH, Nilaparvata lugens Stål) is the most devastating pest of rice in Asia and causes significant yield loss annually. Around 37 BPH resistance genes have been identified so far in indica, African rice varieties along with wild germplasms such as Oryza officinalis, O. minuta, O. nivara, O. punctata, O. rufipogon and O. latifolia. Genes/QTLs involved in BPH resistance, including Bph1, bph2/BPH26, Bph3, Bph6, bph7, BPH9, Bph12, Bph14, Bph15, Bph17, BPH18, bph19, Bph20, Bph21(t), Bph27, Bph27(t), Bph28(t), BPH29, QBph3, QBph4, QBph4.2, Bph30, Bph32, Bph33, Bph35 and Bph36, have been fine-mapped by different researchers across the globe. The majority of genes/QTLs are located on rice chromosomes 1, 3, 4, 6, 11 and 12. Rice plants respond to BPH attack by releasing various endogenous metabolites like proteinase inhibitors, callose, secondary metabolites (terpenes, alkaloids, flavonoid, etc.) and volatile compounds. Besides that, hormonal signal pathways mediating (antagonistic/synergistic) resistance responses in rice have been well studied. Marker-assisted breeding and genome editing techniques can also be adopted for improving resistance to novel BPH biotypes.     

穗发芽率和发芽指数及STS标记Vp1B3在小麦抗穗发芽基因型鉴定中的应用

为了筛选抗穗发芽品种(系)并了解其抗性机制,应用整穗发芽法和发芽指数鉴定了33份小麦新品系的穗发芽抗性,以红粒品种京冬8号和京9428作为抗穗发芽对照,以白粒品种京411和中优9507作为感穗发芽对照,并结合共显性STS标记Vp1B3对其基因型进行检测.结果表明,穗发芽率(SGR)低于抗性对照京冬8号和京9428平均值(12.7%)的品系有7份,其中2份为白粒,5份为红粒.只有一份红粒品系CA0489的发芽指数(GI)低于抗性对照平均值(13.2%).应用STS标记Vp1B3共扩增出849 bp、569 bp和652 bp三种片段,分别属于抗穗发芽基因型Vp1Bb和Vp1Bc及感穗发芽基因型Vp1Ba,其频率分别为3%、18%和79%.红粒抗穗发芽品系CA0489属于Vp1Bb基因型和红色种皮休眠型,CA0481属于Vp1Bc抗穗发芽基因型,另外三份红粒抗穗发芽品系的抗性机理还需要进一步研究;白粒抗穗发芽品系CA0509和CA0459的抗性为非Vp1B3类型,其抗性可能与穗部性状和颖壳抑制物有关.     

Identification of Major Locus Bph35 Resistance to Brown Planthopper in Rice

An introgression line RBPH660, derived from wild rice Oryza rufipogon, showed stable resistance to brown planthopper(BPH). Segregation analysis indicated BPH resistance of RBPH660 was controlled by multiple genes/QTLs. By using the bulked segregant analysis(BSA)-seq method, two genomic regions harboring QTLs resistance to BPH were identified from 1.20 to 16.70 Mb on chromosome 4 and from 10.20 to 12.60 Mb on chromosome 9 in RBPH660, respectively. A major resistance locus, designated as Bph35 accounting for 51.27% of the phenotypic variation with a LOD score of 42.51, was mapped to the candidate region of chromosome 4 between In Del(insertion-deletion) markers PSM16 and R4 M13. For fine mapping of Bph35, one simple sequence repeat and three newly developed In Del markers were used to screen the recombinants. Finally, the Bph35 locus was delimited in the region from 6.28 to 6.93 Mb and there were 18 predicted protein-encoding genes with a total of 114 non-synonymous single nucleotide polymorphism(SNP) variant sites between the resistant and susceptible parents. Out of these genes, Os04 g0193950, encoding a putative NB-ARC(nucleotidebinding adaptor shared by APAF-1, R proteins and CED-4) and LRR(leucine-rich repeat) domain protein with nine non-synonymous SNP substitutions in its coding sequence regions, might be the candidate gene for Bph35. These findings would facilitate the map-based cloning of the Bph35 gene and development of resistant varieties against BPH in rice.     

抗白叶枯病杂交水稻的分子标记辅助育种

应用分子标记辅助回交,育成了带广谱抗白叶枯病基因[i]Xa-21[/i]的两个杂交水稻恢复系R8006和R1176,所配的杂交水稻组合中优6号、中优1176在中国南方稻区及多个省级区试中表现抗病、优质、高产,具有较广的商业开发潜力。