含白叶枯病抗性基因Xa23水稻恢复系的分子标记辅助选育

 通过常规杂交育种技术和分子标记辅助选择技术,对携有白叶枯病抗性基因Xa23的亲本材料CBB23进行回交转育和农艺性状改良。通过分子标记辅助选择检测目的基因,快速获得了12份具有Xa23基因的恢复系稳定材料。采用白叶枯病菌株的专化强毒菌系P6对该12份材料进行室内接种鉴定,筛选出1份高抗白叶枯病的抗性新恢复系材料。应用3个不同的不育系对该恢复系进行测交,结果表明该恢复系具有良好的恢复性及产量潜力。     

四川主要水稻恢复系抗白叶枯病基因的分子检测

用分别与水稻抗白叶枯病基因Xa4、Xa7、Xa21和Xa23紧密连锁的PCR标记MP12、M3、pTA248和RM206对四川省新选育的以及主要应用的39份恢复系进行了分子检测，结果表明，这些恢复系均不含有Xa7（M3）和Xa21（pTA248）抗性等位点，17份携有榭（MP12）抗性等位点，少数（6个）携有Xa23（RM206）抗性等位点。在杂交水稻恢复系抗性改良中，应采用分子标记辅助选择（MAS）加快引入利用除工卅外的其它儿个显性抗性基因。     

利用基因组文库加速Xa23基因定位的染色体步移

 用距离水稻抗白叶枯病基因Xa23 0.8 cM的EST标记C189，扩增Xa23的近等基因系CBB23的基因组片段（0.8 kb）为探针,筛选水稻明恢63的TAC文库和广陆矮4号的PAC文库，对获得的7个阳性克隆用酶切法和Tail PCR法进行末端片段分离，获得15个末端片段， 用于感病亲本金刚30和抗病亲本CBB23之间的多态性检测，发现7个末端片段在双亲间有多态性，分别为69B、70N、81N、45B、45N、84N和84B。用这些末端片段作RFLP标记，对金刚30/CBB23的F2群体进行检测和连锁分析,结果表明这些标记与Xa23的遗传距离依次为0.4、0.4、0.4、0.5、0.6、0.6和1.1 cM。虽然69B、70N和81N与Xa23的遗传距离均为0.4 cM，但序列比对揭示69B与70N的物理距离为35 kb，与81N为95 kb，69B距Xa23最近。三者与Xa23的遗传距离虽然相同，但物理距离存在很大差异。这些末端片段标记加密了Xa23基因一侧的遗传图谱，并使其遗传距离缩短到0.4 cM，加速了Xa23的定位进程，为Xa23的分离克隆奠定了重要基础。讨论了这种染色体步移方法的适用条件。     

疣粒野生稻抗白叶枯病新基因的初步鉴定

以10个菲律宾白叶枯病菌小种和1个中国白叶枯病菌小种为供试菌系,以高感白叶枯病水稻品种IR24及携有不同抗白叶枯病基因的近等基因系IRBB1等16个材料作为参照,对粳稻品种8411/疣粒野生稻体细胞杂交获得的两个抗白叶枯病新种质SH5和SH76进行了白叶枯病抗谱鉴定。结果表明SH5和SH76在苗期的抗谱较广,并且与已知抗病基因的抗谱不同,但与IRBB5(xa5)和IRBB7(Xa7)相似。分别用xa5和Xa7的分子标记2F_1R和M5进行检测,确定SH5和SH76中不含有xa5和Xa7基因。初步推测SH5、SH76可能含有一个新的抗病基因或者一个连锁的基因簇群。     

转基因杂交稻组合汕优63和汕优559对白叶枯病的抗性

 采用包括与Xa21相匹配的鉴别菌系在内的12个国际鉴别小种和中国7个病原型，接种由明恢63、盐恢559的Xa21转基因单拷贝抗性纯合系与珍汕97A杂交获得的转基因杂交稻汕优63 (汕优63-Xa21 73个 F1群体和转基因杂交稻汕优559（汕优559-Xa21 13个F1群体，测定转基因杂交稻对白叶枯病的抗性。结果表明这2个转基因杂交稻组合具有优良的广谱抗性， 抗谱与Xa21基因供体IRBB21相同，非转基因杂交稻表现感病。     

抗白叶枯病水稻不育系先抗A和天抗A的选育

分别以保持先B和天B为受体和轮回亲本,以携带抗白叶枯病基因Xa23的育种材料P59为供体,通过杂交、回交和分子标记辅助选择,结合农艺性状表现和田间接种抗性鉴定,育成2个含有Xa23基因且抗白叶枯病的保持系先抗B和天抗B,再分别与先A与天A测交、成对回交,育成抗白叶枯病水稻不育系先抗A和天抗A,于2013年9月通过广西科技厅田间技术鉴定。这2个不育系具有败育彻底、农艺性状优良、异交结实率高和抗白叶枯病等特点,配制的杂交稻组合表现出明显的白叶枯病抗性和产量优势,具有较好的应用前景。     

分子标记辅助选择改良Ⅱ-32B白叶枯病抗性

水稻白叶枯病是危害水稻生产的重要病害之一。以含抗水稻白叶枯病Xa23基因的IRBBL23为抗性供体,以感病Ⅱ-32B为轮回亲本,采用与Xa23基因紧密连锁标记CP02662检测后代单株。结果表明：通过田间抗性鉴定和分子标记辅助选择相结合的方法,最终获得6个导入Xa23基因白叶枯病抗性单株,病斑长度为0.34~1.34 cm。利用均匀分布水稻染色体上的178对SSR引物,对入选单株的背景恢复率进行分析。结果表明,6个入选单株的背景恢复率为96％~100％,基本恢复为轮回亲本的遗传背景。     

利用分子标记辅助育种技术选育高抗白叶枯病恢复系

通过常规回交育种结合分子标记辅助选择技术,已将来自IRBB24的2个抗白叶枯病基因, Xa21和Xa4, 聚合到感病的杂交稻恢复系绵恢７２５中。通过分子标记检测目标基因和亲本遗传背景差异分析快速获得了4个高抗白叶枯病的恢复系姊妹系R207 1、R207 2、R207 3和R207 4。采用与目标抗性基因相应的菲律宾菌系P1、P6 和7个中国病原型代表菌系（CⅠ~CⅦ）对4个姊妹系及其杂交组合进行田间接种。结果表明,这些姊妹系及所配制的杂交组合抗病性强、抗谱广,其中杂交组合G46A/R207 2具有良好的产量潜力,将R207 2定名为蜀恢207。     

优质抗稻瘟病水稻三系不育系M20A的选育

以改良优质三系不育系金山A-1的稻瘟病抗性和配合力作为育种目标,利用携有抗稻瘟病基因Pi-9的金23B抗病近等基因系(04AMA-88)与携有抗白叶枯病基因Xa21的金山B-1抗病近等基因系(04AMA-49)杂交,经分子标记辅助选择,在杂交F4代中选择携带Pi-9的优良单株与金23A测交并转育不育系,育成抗稻瘟病、米质优、配合力好、异交率高的新不育系M20A。该不育系及其配制的杂交早稻新组合M优2155(M20A/明恢2155)于2014年5月通过福建省品种审定委员会审定。     

利用分子标记辅助选择改良温恢117白叶枯病抗性

以恢复系温恢117为受体,以IRBB21为抗白叶枯病基因Xa21的供体,通过杂交、多次回交和自交,结合分子标记辅助选择技术,将Xa21基因导入温恢117中,结合抗性基因Xa21的分子检测和农艺性状表现,筛选出带有Xa21基因的15个恢复系株系.采用白叶枯病菌株的专化强毒菌系P6进行接种鉴定,各株系的白叶枯病抗性均强于受体亲本温恢117,其中株系M033抗性与IRBB21相当,而农艺性状与温恢117相近.     

Improvement of bacterial blight resistance of hybrid rice in China using the Xa23 gene derived from wild rice (Oryza rufipogon)

A novel bacterial blight (BB) resistance gene, Xa23, identified from Oryza rufipogon was introgressed into three popular restorer lines (Minghui63, YR293 and Y1671) for wild abortive cytoplasmic male sterility by marker-assisted backcross breeding approach in combination with artificial inoculation and stringent phenotypic selections. The three derived BB resistant restorer lines (Minghui63-Xa23, YR293-Xa23 and Y1671-Xa23) and their hybrid combinations with Zhenshan97A (Shanyou63-Xa23), NongfengA (Fengyou293-Xa23) and Zhong9A (Zhongyou1671-Xa23) demonstrated similar BB resistance spectrum as the donor parent, CBB23 (B). The newly developed BB resistant restorers and their derived hybrids were identical to their respective original versions for agronomic traits especially under disease free condition. However, under severe disease condition, the three BB resistant restorer lines exhibited significantly higher grain weight and spikelet fertility as compared to the respective original restorer lines thus further resulting in BB resistant hybrids with significantly higher grain yields than their respective popular original hybrids. The results indicated that the Xa23 gene could completely express its dominant broad spectrum resistance in different backgrounds of both restorer and male sterile lines across different growth stages, suggesting its immense breeding value in BB resistance improvement for hybrid rice. Moreover, a reasonable utilization and deployment of Xa23 gene for efficient control of BB disease in hybrid rice production was recommended.     

利用功能标记鉴定普通野生稻中的白叶枯病抗性基因

 以9个菲律宾白叶枯病菌小种对供试的9份普通野生稻（Oryza rufipogon Griff.）及1份高感白叶枯病材料IR24进行抗性鉴定,发现IR24对所有的菌株都表现为高感, 6份野生稻材料对9个菌株表现全抗,占参试野生稻总数的67%。取自广西玉林的1份材料只感PXO280(P8),海南万宁的1份材料感PXO71(P4),广州高州的1份材料对PXO79、PXO99和PXO339感病,而这几份材料对其余菌株都表现为抗病,说明每份材料至少含有1个抗性基因。利用已克隆的白叶枯病抗性基因xa5、xa13、Xa21和Xa27的功能标记检测,结果表明9份供试普通野生稻中都不含抗性基因xa5、Xa21;5份为显性Xa13纯合体,4份为隐性抗病xa13杂合体;5份为抗病显性Xa27纯合体,3份为隐性xa27纯合体,1份材料中xa27和Xa27都不存在。     

Application of Functional Markers to Identify Genes for Bacterial Blight Resistance in Oryza rufipogon

Field resistances of nine accessions of common wild rice (Oryza rufipogon Griff.) and one rice variety (IR24) were evaluated by using nine strains of bacterial blight pathogen (Xanthomonas oryzae pv. oryzae) from the Philippines. IR24 was highly susceptible to all the strains, and six common wild rice accessions resisted all the nine strains, with a resistance frequency of 67%. The accessions Yulin and Wanning were only susceptible to PXO280 and PXO71, respectively. The accession Gaozhou was susceptible to the three strains PXO79, PXO99 and PXO339, whereas resistant to the other six strains. It could be concluded that there is at least one resistance gene in each common wild rice accession. The functional markers of the genes xa5, xa13, Xa21 and Xa27 were used to detect the presence of these resistance genes in the nine tested wild rice accessions, and it was found that four wild rice accessions contained heterozygous xa13. Among the nine common wild rice accessions, five were homozygous for Xa27 and three homozygous for xa27, and the accession Laibin contained neither xa27 nor Xa27. In addition, there were no xa5 and Xa21 in all of these accessions.     

Marker-Assisted Selection of Xa21 Conferring Resistance to Bacterial Leaf Blight in indica Rice Cultivar LT2

Bacterial leaf blight of rice (BLB), caused by Xanthomonas oryzae pv. oryzae, is one of the most destructive diseases in Asian rice fields. A high-quality rice variety, LT2, was used as the recipient parent. IRBB21, which carries the Xa21 gene, was used as the donor parent. The resistance gene Xa21 was introduced into LT2 by marker-assisted backcrossing. Three Xoo races were used to inoculate the improved lines following the clipping method. Eleven BC₃F₃ lines carrying Xa21 were obtained based on molecular markers and agronomic performance. The 11 lines were then inoculated with the three Xoo races. All the 11 improved lines showed better resistance to BLB than the recipient parent LT2. Based on the level of resistance to BLB and their agronomic performance, five lines (BC₃F₃ 5.1.5.1, BC₃F₃ 5.1.5.12, BC₃F₃ 8.5.6.44, BC₃F₃ 9.5.4.1 and BC₃F₃ 9.5.4.23) were selected as the most promising for commercial release. These improved lines could contribute to rice production in terms of food security.