Use of Major Quantitative Trait Loci to Improve Grain Yield of Rice

Further improvement of rice productivity remains a challenge. Breeding is perceived as an important option to increase rice yield. However, the genetic progress of grain yield in most rice breeding programs was slow in the last decades. Although great progress in rice genomics and molecular biology has been achieved, the effect of such technological innovations on rice breeding is far small. Marker-assisted selection （MAS） for a few target quantitative trait loci （QTLs） has significant effects in improving qualitative traits, such as disease resistance. The success of MAS has therefore motivated breeders to identify and use major QTLs for yield and yield component traits. In this review, we summarized the recent methods in QTL identification, including novel statistical methods for linkage and association mapping, special population types, and whole-genome sequencing. We reviewed the successful application of marker-assisted gene introgression and gene pyramiding to improve grain yield and discussed the design of efficient MAS schemes to further increase the success rate of breeding programs. The use of well-characterized major QTLs through introgression and gene pyramiding is proven effective in improving grain yield, particularly yield under abiotic stress. Major QTLs that are stable across genetic background and growing environments are often found in less adapted germplasms, such as landraces and wild relatives. Advanced backcross QTL analysis and introgression lines, which integrate QTL discovery and utilization, are important methods for exploiting major QTLs contained in such germplasms. Next-generation sequencing substantially increases mapping resolution and accelerates the identification of casual genes underlying major QTLs. Practical guidelines derived from theoretical and empirical studies are given to guide the design of efficient marker-assisted gene introgression and pyramiding schemes.     

水稻耐盐基因定位与克隆及品种耐盐性分子标记辅助选择改良研究进展

土壤盐渍化严重制约水稻生产发展,提高耐盐性已成为水稻育种的重要目标之一。挖掘水稻耐盐新基因,解析其分子作用机制可以为水稻耐盐性遗传改良奠定基础。本文从定位群体、耐盐性鉴定时期和鉴定方法、耐盐性评价指标、鉴定到的耐盐QTL、耐盐QTL的精细定位和图位克隆等方面,总结了近年来水稻耐盐QTL定位研究中所取得的进展;介绍了水稻耐盐/盐敏感突变体筛选和基因克隆以及耐盐性关联分析的研究近况;并对水稻耐盐性分子标记辅助选择改良的现状作了概述。     

Lowering Grain Amylose Content in Backcross Offsprings of indica Rice Variety 057 by Molecular Marker-Assisted Selection

To lower the amylose content （AC） of the indica rice restorer line 057 with high AC, backcrosses were made respectively by using four indica varieties （R367, 91499, Yanhui 559, Hui 527） as low AC donor parents and 057 as the recurrent parent. A molecular marker （PCR-Acc Ⅰ） was used to identify the genotypes （GG, TT and GT） of the waxy （Wx） gene. Plants with GT genotype were selected and used as female parent and crossed with 057 to advance generation. The ACs of rice grains harvested from plants with different Wx genotypes were measured and compared to analyze the efficiency of marker-assisted selection. The ACs of the rice grain, harvested from the plants of Wx genotypes GG, GT and TT, were higher than 20%, in the range of 17.7-28.5%, and less than 18%, respectively. The PCR-Acc Ⅰ marker could be used for efficiently lowering the AC of 057 through backcrossing, and there were some influence of parental genetic background on the AC of rice grains with the same Wx genotype.     

Functional Marker Development and Effect Analysis of Grain Size Gene GW2 in Extreme Grain Size Germplasm in Rice

GW2 is an important gene that regulates grain width and weight. We used c DNA clone to obtain the sequences of GW2 from large- and small-grained rice varieties,TD70 and Kasalath,respectively. Then,we developed a d CAPS(derived cleaved amplified polymorphic sequence) marker on the basis of the sequence difference between functional and nonfunctional GW2 genes to analyze the genotypes and phenotypes of recombinant inbred lines. Results showed that the sequence of GW2TD70 had a single nucleotide deletion at site 316 that generates a termination codon. This codon terminated the GW2 protein in advance. By contrast,the sequence of GW2 Kasalath encoded an intact protein. A novel d CAPS marker was designed in accordance with a base A deletion at site 316 of the sequence. After the PCR product was digested by Apo I,TD70 showed 21 and 30 bp fragments,and Kasalath showed a 51 bp fragment. Up to 82 lines contained GW2TD70,and 158 lines contained GW2 Kasalath. The lines that contained TD70 alleles displayed substantial increases in width and 1000-grain weight. This result suggested that GW2 played a critical role in rice breeding.     

Transferring Translucent Endosperm Mutant Gene Wx-mq and Rice Stripe Disease Resistance Gene Stv-bi by Marker-Assisted Selection in Rice (Oryza sativa)

A high-yielding japonica rice variety, Wuyunjing 7, bred in Jiangsu Province, China as a female parent was crossed with a Japanese rice variety Kantou 194, which carries a rice stripe disease resistance gene Stv-bi and a translucent endosperm mutant gene Wx-mq. From F2 generations, a sequence characterized amplified region (SCAR) marker tightly linked with Stv-bi and a cleaved amplified polymorphic sequence (CAPS) marker for Wx-mq were used for marker-assisted selection. Finally, a new japonica rice line, N...     

通过分子标记辅助选择技术选育携有水稻白叶枯病抗性基因Xa23的水稻株系

 通过杂交和分子标记辅助选择技术,将亲本材料18113携有的水稻白叶枯病抗病基因Xa23导入亚种间恢复系H705和优质恢复系H706。利用EST标记C189检测目的基因,在18113/H705 F3和18113/H706 F3株系中,分别获得71和52份携有Xa23纯合基因型恢复系。采用水稻白叶枯病Ⅳ型小种代表菌株浙173进行剪叶接种,分别鉴定出61和44份抗病株系,它们的分子标记辅助选择准确率分别为85.92%和84.61%。研究结果表明,C189是检测Xa23基因的有效分子标记之一,不过,其假阳性的几率高于理论交换率的现象还有待于进一步研究。     

分子标记辅助选择抗稻白叶枯病基因在选育水稻广亲和恢复系上的应用

 以IRBB21为Xa21基因供体,广亲和恢复系4183为受体，进行1次杂交并回交3次，逐代用分子标记检测手段，导入广谱抗白叶枯病基因Xa21,在保持广亲和恢复系4183优良经济性状的基础上，增强其抗白叶枯病的能力，育成抗白叶枯病的广亲和恢复系抗4183，其抗性达到了IRBB21的抗性水平，且保持了4183的广亲和性、恢复性及优良的经济性状。并就杂交水稻白叶枯病的抗性改良及育种对策进行了讨论。     

Breeding Rice Restorer Lines with High Resistance to Bacterial Blight by Using Molecular Marker-Assisted Selection

Rice bacterial blight, one of the major diseases of rice caused by Xanthomonas oryzae pv. oryzae, Xoo, jeopardizes rice diversely. It causes leaf wilting, affects photosynthesis and reduces 1000-grain weight and generally results in yield loss by 20-30 pe…     

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。     

Transferring Translucent Endosperm Mutant Gene Wx-mq and Rice Stripe Disease Resistance Gene Stv-b by Marker-Assisted Selection in Rice (Oryza sativa)

A high-yielding japonica rice variety, Wuyunjing 7, bred in Jiangsu Province, China as a female parent was crossed with a Japanese rice variety Kantou 194, which carries a rice stripe disease resistance gene Stv-bⁱ and a translucent endosperm mutant gene Wx-mq. From F₂ generations, a sequence characterized amplified region (SCAR) marker tightly linked with Stv-bⁱ and a cleaved amplified polymorphic sequence (CAPS) marker for Wx-mq were used for marker-assisted selection. Finally, a new japonica rice line, Ning 9108, with excellent agronomic traits was obtained by multi-generational selection on stripe disease resistance and endosperm appearance. The utilization of the markers from genes related to rice quality and disease resistance was helpful not only for establishing a marker-assisted selection system of high-quality and disease resistance for rice but also for providing important intermediate materials and rapid selection method for good quality, disease resistance and high yield in rice breeding.     

一种水稻香味基因功能标记的开发

为了提高水稻香味基因的分子标记辅助选择的准确性,根据水稻隐性香型品种与非香型品种的BAD2基因（即香味基因）序列之间存在8 bp碱基缺失,设计出香味基因fgr的InDel功能标记GRFM04。利用该标记对粤丰B（香型）/振丰B（非香型）的F2分离群体和其他16份香稻品种和6份非香稻品种进行检测验证。依据其PCR扩增产物电泳带型,可以准确地区分出香型纯合基因型、非香型纯合基因型和杂合基因型3种带型,且3种带型与其植株或品种相应的香味性状表现完全呈一一对应关系。     

分子标记辅助选择聚合水稻暗胚乳突变基因Wx-mq和抗条纹叶枯病基因Stv-b~i

以江苏高产粳稻品种武运粳7号为母本,具有条纹叶枯病抗性基因Stv-bi和暗胚乳突变基因Wx-mq的日本粳稻品种关东194作父本配制杂交组合进行聚合育种。利用与Stv-bi基因共分离的SCAR标记及与暗胚乳突变基因Wx-mq紧密连锁的CAPS功能标记对其分离世代进行标记位点的检测,结合田间多代选育、抗性鉴定和籽粒胚乳外观鉴定,将条纹叶枯病抗性基因Stv-bi和暗胚乳突变基因Wx-mq同时转育到高产品种中,筛选、培育出优质、抗病、高产且农艺性状优良的水稻新品系宁9108。利用与水稻优质及抗病基因紧密连锁的分子标记,不仅初步建立了优质、抗病育种的分子标记辅助选择体系,也为水稻优质、抗病、高产育种提供了一种简单、快捷的选择方法和重要的中间材料。     

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.     

利用分子标记技术降低协优57的直链淀粉含量

 为克服产量高、适应性广的中籼组合协优57的直链淀粉含量较高、蒸煮食味品质较差的缺点,对其父母本较高的直链淀粉含量特性进行了改良。先前利用PCR AccⅠ分子标记辅助选择对其父本057进行改良,育成了直链淀粉含量改良型057, 简称057(TT)。本研究以ND42为优质基因的供体,利用PCR AccⅠ分子标记对协青早B\[母本协青早A的保持系,简称协青早B(GG)\]进行改良,得到直链淀粉含量改良型协青早B,简称协青早B(TT)。然后用改良前、后的各亲本分别配组,分析各组合的基因型(GG、GT、TT)和食味品质。结果表明,改良单亲的GT型组合协青早B(GG)/057(TT)、协青早B(TT)/057(GG)杂交稻米的直链淀粉含量由原组合协青早B(GG)/057(GG)的26.1%分别降到19.3%和19.2%,但均一性较差。改良双亲的TT纯合型组合协青早B(TT)/057(TT)的杂交稻米,不仅直链淀粉含量降到中等偏低水平(12.5%)、胶稠度变得更软,而且直链淀粉含量的均一性也有了很大的提高。     

分子标记辅助选择降低籼稻057的直链淀粉含量

 以4种低直链淀粉含量（AC）的籼稻品种(R367、91499、盐恢559、恢527)作优质基因的供体，以产量配合力较强的三系籼稻恢复系057为受体轮回亲本，对高AC值的057进行回交改良。在回交改良中利用分子标记对控制AC值的基因型进行选择，并对分子标记鉴定的Wx基因3种表达类型（GG、TT、GT）植株稻米的AC值进行测定分析。结果显示，GG型植株的稻米AC值高于20%；GT型的变化较大，为17.71%~28.48%；TT型的小于18%。经最小显著差数法（LSR）检验，3种基因型间AC差异达极显著水平。表明这种分子标记对稻米AC值的辅助选择是可靠有效的。通过分子标记辅助选择有效降低了057的AC值。     

水稻基因设计育种的研究进展与展望

 分子标记、基因工程（转基因技术）和基因组学的研究成果为实现对基因型的直接选择和农作物新品种设计育种提供了可能。设计育种旨在控制所有重要农艺性状基因的所有等位性变异。通过精细遗传定位、高饱和染色体片段系的构建和表型鉴定,可以实现设计育种。鉴于一系列可以得到的标记技术、软件工具以及已定位和克隆的水稻基因、已建立的基因 表型（G P）数据关联信息,设计育种目标已成为现实。主要介绍了设计育种、分子育种及一些基本技术,当前重点应用的基因功能研究与水稻功能基因分子利用的研究进展,并对水稻基因设计育种的前景进行了展望。     

Molecular Scree ning of Blast Resistance Genes in Rice Germplasms Resistant to Magnaporthe oryzae

Molecular screening of major rice blast resistance genes was determined with molecular markers, which showed close-set linkage to 11 major rice blast resistance genes (Pi-d2, Pi-z, Piz-t, Pi-9, Pi-36, Pi-37, Pi5, Pi-b, Pik-p, Pik-h and Pi-ta²), in a collection of 32 accessions resistant to Magnaporthe oryzae. Out of the 32 accessions, the Pi-d2 and Pi-z appeared to be omnipresent and gave positive express. As the second dominant, Pi-b and Piz-t gene frequencies were 96.9% and 87.5%. And Pik-h and Pik-p gene frequencies were 43.8% and 28.1%, respectively. The molecular marker linkage to Pi-ta² produced positive bands in eleven accessions, while the molecular marker linkage to Pi-36 and Pi-37 in only three and four accessions, respectively. The natural field evaluation analysis showed that 30 of the 32 accessions were resistant, one was moderately resistant and one was susceptible. Infection types were negatively correlated with the genotype scores of Pi-9, Pi5, Pi-b, Pi-ta² and Pik-p, although the correlation coefficients were very little. These results are useful in identification and incorporation of functional resistance genes from these germplasms into elite cultivars through marker-assisted selection for improved blast resistance in China and worldwide.     

水稻粒重及其相关性状的遗传解析

 应用292个 Lemont/ 特青F13重组自交系(RILs)和272个标记的遗传连锁图谱分析粒重及籽粒长、宽、厚、长/宽、体积和容重6个相关性状的遗传。所有性状在RILs中出现超亲分离。粒重与其他籽粒性状均呈显著正相关，而涉及籽粒品质的容重与粒宽、粒厚呈显著负相关。检测到影响粒重及其相关性状的主效QTL 48个和互作位点38对，这些QTL解释了各性状55%以上的表型变异。80%以上的主效QTL呈现一因多效或生理相关。QGl3和 QGl2等通过籽粒体积间接影响粒重， Lemont增效等位基因有利于籽粒品质的改良，而QGw5、QGt1和QGv7的特青增效等位基因增加粒重同时降低籽粒容重及品质。通过标记辅助选择进行不同QTL的重组，有望在增加粒重的同时改善籽粒品质。