Genetic analysis and molecular mapping of a novel gene for zebra mutation in rice （Oryza sativa L.）

A novel zebra mutant, zebra-15, derived from the restorer line JinhuilO （Oryza sativa L. ssp. indica） treated by EMS, displayed a distinctive zebra leaf from seedling stage to jointing stage. Its chlorophyll content decreased （55.4%） and the ratio of Chla/Chlb increased （90.2%） significantly in the yellow part of the zebra-15, compared with the wild type. Net photosynthetic rate and fluorescence kinetic parameters showed that the decrease of chlorophyll content significantly influenced the photosynthetic efficiency of the mutant. Genetic analysis of F2 segregation populations derived from the cross of XinonglA and zebra-15 indicated that the zebra leaf trait is controlled by a single recessive nuclear gene. Ninety-eight out of four hundred and eighty pairs of SSR markers showed the diversity between the XinonglA and the zebra-15, their F2 population was then used for gene mapping. Zebra-15 （Z-15） gene was primarily restricted on the short arm of chromosome 5 by 150 F2 recessive individuals, 19.6 cM from marker RM3322 and 6.0 cM from marker RM6082. Thirty-six SSR markers were newly designed in the restricted location, and the Z-15 was finally located between markers nSSR516 and nSSR502 with the physical region 258 kb by using 1,054 F2 recessive individuals.     

Molecular evolution and functional divergence of HAK potassium transporter gene family in rice （Oryza sativa L.）

The high-affinity K＋ （HAK） transporter gene family is the largest family in plant that functions as potassium transporter and is important for various aspects of plant life. In the present study, we identified 27 members of this family in rice genome. The phylogenetic tree divided the land plant HAK transporter proteins into 6 distinct groups. Although the main characteristic of this family was established before the origin of seed plants, they also showed some differences between the members of non-seed and seed plants. The HAK genes in rice were found to have expanded in lineage-specific manner after the split of monocots and dicots, and both segmental duplication events and tandem duplication events contributed to the expansion of this family. Functional divergence analysis for this family provided statistical evidence for shifted evolutionary rate after gene duplication. Further analysis indicated that both point mutant with positive selection and gene conversion events contributed to the evolution of this family in rice.     

Identification of quantitative trait loci for four morphologic traits under water stress in rice （Oryza sativa L.）

Late season drought coinciding with the rice booting to heading stage affects the development of plant height,panicle exsertion,and flag leaf size,and causes significant yield loss.In this study,a recombinant inbred line population derived from a cross between paddy and upland cultivars was used for data collection of the morphologic traits under well water and drought stress conditions.bought stress was applied at the stage of panicle initiation in the field in 2002 and at the booting stage in PVC pipes in 2003.The data from stress con ditions and their ratios(tait measured under stress condition/trait measured under well water condition)or differences(trait measured under stress condition minus trait measured under well water condition)were used for OTL analysis.Totally,17 and 36 QTLs for these traits were identified in 2002 and 2003,respectively,which explained a range of 2.58%-29.82%Of the phenotypic variation.Among them,six QTLs were commonly identified in the two years,suggesting that the drought stress in the two years was different.The genetic basis of these traits will provide useful information for improving rice late season drought resistance,and their application as indirect indices in rice late season drought resistance screening was also discussed.     

Additive and additive x additive interaction make important contributions to spikelets per panicle in rice near isogenic （Oryza sativa L.） lines

Epistasis plays an important role in the genetic basis of rice yield traits. Taking interactions into account in breeding programs will help the development of high-yielding rice varieties. In this study, three sets of near isogenic lines (NILs) targeting three QTLs for spikelets per panicle (SPP), namely qSPP1, qSPP2 and qSPP7, which share the same Zhenshan 97 genetic background, were used to produce an F₂ population in which the three QTLs segregated simultaneously. The genotypes of the individual F₂ plants at the three QTLs were replaced with three markers that are closely linked to the corresponding QTLs. These QTLs were validated in the F₂ and F₃ populations at the single marker level. qSPP7 exhibited major pleiotropic effects on SPP, plant height and heading date. Multifactor analysis of variance was performed for the F₂ population and its progeny. Additive (additive interaction between qSPP2 and qSPP7 had significant effects on SPP in both the F₂ population and its progeny. Both additive and additive (additive interactions could explain about 73% of the total SPP phenotypic variance. The SPP performance of 27 three-locus combinations was ranked and favorable combinations were recommended for rice breeding in different ecosystems.     

Introgression of qPE9-1 allele, conferring the panicle erectness, leads to the decrease of grain yield per plant in japonica rice （Oryza sativa L.）

Panicle architecture is closely related to yield formation. The qPE9-1 gene has been proved to be widely used in high-yield rice cultivar developments, conferring erect panicle character in japonica rice. Recently, qPE9-1 has been successfully cloned; however, the genetic effect on grain yield per plant of the erect panicle allele qPE9-1 is controversial yet. In the present study, a drooping panicle parent Nongken 57, carrying qpe9-1 allele, was used as recurrent parent to successively backcross to a typical erect panicle line from the double haploid (DH) population (Wuyunjing 8/Nongken 57), which was previously shown to carry qPE9-1 allele. Thus a pair of near-isogenic lines (NILs) was developed. The comparison of agronomic traits between the NILs showed that, when qpe9-1 was replaced by qPE9-1, the panicle architecture was changed from drooping to erect; moreover, the panicle length, plant height, 1000-grain weight and the tillers were significantly decreased, consequently resulting in the dramatic decrease of grain yield per plant by 30%. Therefore, we concluded that the qPE9-1 was a major factor controlling panicle architecture, and qPE9-1 had pleiotropic nature, with negative effects on grain yield per plant. This result strongly suggests that the erect panicle allele qPE9-1 should be used together with other favorable genes in the high-yield breeding practice. In addition, the effect of qPE9-1 on eating and cooking quality was also discussed in the present study.     

不同遗传背景及环境中水稻（Oryza sativa L.）穗长的QTLs和上位性分析

以粳稻Ａｚｕｃｅｎａ为父本与灿稻ＩＲ６４杂交发展的一双单倍体（ＤＨ） 本,与灿稻ＩＲ１５５２杂交发展的一重组自交系（ＲＩ）群体为材料,应用分子标记图说对２个群体在大田答舅栽２个环境下的穗长进行ＱＴＬｓ及上位性效应分析,ＤＨ群体中共检测６个穗长ＱＴＬｓ,位于第１、４长染色体上的３个ＱＴＬｓ,,在２个环境中稳定表达,未检测一闰性效应,加性效应为穗长遗传主效应,ＲＩ群体中,共检测到３个穗长ＱＴＬｓ及６对     

Expression of PIN Genes in Rice （Oryza sativa L.）： Tissue Specificity and Regulation by Hormones

Twelve genes of the PIN family in rice were analyzed for gene and protein structures and an evolutionary relationship with reported AtPINs in Arabidopsis. Four members of PIN1 （designated as OsPINla-d）, one gene paired with AtPIN2 （OsPIN2）, three members of PIN5 （OsPIN5a-c）, one gene paired with AtPIN8 （OsPIN8）, and three monocot-specific PINs （OsPIN9, OsPINIOa, and b） were identified from the phylogenetic analysis. Tissue-specific expression patterns of nine PIN genes among them were investigated using RT-PCR and GUS reporter. The wide variations in the expression domain in different tissues of the PIN genes were observed. In general, PIN genes are up-regulated by exogenous auxin, while different responses of different PIN genes to other hormones were found.     

Subspecies-specific intron length polymorphism markers reveal clear genetic differentiation in common wild rice （Oryza rufipogon L.） in relation to the domestication of cultivated rice （O. sativa L.）

It is generally accepted that Oryza rufipogon is the progenitor of Asian cultivated rice （O. sativa）. However, how the two subspecies of O. sativa （indica and japonica） were domesticated has long been debated. To investigate the genetic differentiation in O. rufipogon in relation to the domestication of O. sativa, we developed 57 subspecies-specific intron length polymorphism （SSILP） markers by comparison between 10 indica cultivars and 10 japonica cultivars and defined a standard indica rice and a standard japonica rice based on these SSILP markers. Using these SSILP markers to genotype 73 O. rufipogon accessions, we found that the indica alleles and japonica alleles of the SSILP markers were predominant in the O. rufipogon accessions, suggesting that SSILPs were highly conserved during the evolution of O. sativa. Cluster analysis based on these markers yielded a dendrogram consisting of two distinct groups： one group （Group I） comprises all the O. rufipogon accesions from tropical （South and Southeast） Asia as well as the standard indica rice; the other group （Group II） comprises all the O. rufipogon accessions from Southern China as well as the standard japonica rice. Further analysis showed that the two groups have significantly higher frequencies of indica alleles and japonica alleles, respectively. These results support the hypothesis that indica rice and japonica rice were domesticated from the O. rufipogon of tropical Asia and from that of Southern China, respectively, and suggest that the indica-japonica differentiation should have formed in O. rufipogon long before the beginning of domestication. Furthermore, with an O. glaberrima accession as an outgroup, it is suggested that the indica-japonica differentiation in O. ruffpogon might occur after its speciation from other AA-genome species.     

Mapping QTLs for seed yield and drought susceptibility index in soy-bean（Glycine max L.） across different environments

Drought stress has long been a major constraint in maintaining yield stability of soybean （Glycine max （L.） Merr.） in rainfed ecosystems. The identification of consistent quantitative trait loci （QTL） involving seed yield per plant （YP） and drought susceptibility index （DSI） in a population across different environments would therefore be important in molecular marker-assisted breeding of soybean cultivars suitable for rainfed regions. The YP of a recombinant line population of 184 F2：7：11 lines from a cross of Kefengl and Nannong1138-2 was studied under water-stressed （WS） and well-watered （WW） conditions in field （F） and greenhouse （G） trials, and DSI for yield was calculated in two trials. Nineteen QTLs associated with YP-WS and YP-WW, and 10 QTLs associated with DSI, were identi- fied. Comparison of these QTL locations with previous findings showed that the majority of these regions control one or more traits re- lated to yield and other agronomic traits. One QTL on molecular linkage group （MLG） K for YP-F, and two QTLs on MLG C2 for YP-G, remained constant across different water regimes. The regions on MLG C2 for YP-WW-F and MLG H for YP-WS-F had a pleiotropic effect on DSI-F, and MLG A1 for YP-WS-G had a pleiotropic effect on DSI-G. The identification of consistent QTLs for YP and DSI across different environments will significantly improve the efficiency of selecting for drought tolerance in soybean.     

水稻（Oryza sativa L.）核基因组存在叶绿体psbA基因的同源 …

序列比较说明、重复ＤＮＡ顺序ｐＲＲＤ９＾＊与水稻叶绿体基因组中编码ＱＢ蛋白的ｐｓｂＡ基因存在高度的同源。用ｐＲＢＤ９亚克隆片段ｐＲＲＤ９Ｒ和片段ｐＲＲＤ９Ｌ对水稻的叶绿体和核ＤＮＡ进行Ｓｏｕｔｈｅｒｎ杂交分析,揭示了ｐｓｂＡ基因同源片段在某个进化时期由叶绿体基因组转移到水稻核基因组,而且两者在水稻进化过程中的变异程度存在明显的差异。     

粳稻穗角与稻米品质的相关性及稻米品质遗传分析

测定了粳稻直立穗品种丙8979与弯曲穗品种C堡杂交组合的P1、P2及其重组自交系349个株系的穗角和10个稻米品质性状, 分析了穗角与稻米品质性状之间的相关性, 并运用主基因+多基因混合遗传模型, 对稻米品质10个性状进行了遗传分析。结果表明,穗角与糙米率、整精米率、垩白粒率、垩白度、糊化温度、胶稠度和直链淀粉含量均无显著相关; 与精米率呈显著正相关(r=0.124*); 与粒长和长宽比均呈极显著正相关(相关系数分别为0.470**和0.241**)。糙米率、精米率和直链淀粉含量均受2对主基因+多基因控制, 2对主基因具有累加作用和加性×加性的上位性作用; 整精米率、粒长、长宽比和胶稠度受2对加性-上位性主基因+多基因控制;垩白粒率、垩白度和糊化温度均受3对加性-上位性主基因+多基因控制。糙米率、精米率、整精米率、垩白粒率、垩白度和糊化温度6个品质性状以主基因遗传为主,粒长、长宽比、胶稠度和直链淀粉含量4个性状以多基因遗传为主。     

水稻分蘖角度的QTL定位和主效基因的遗传分析

利用水稻籼粳亚种间组合Asominori×IR24重组自交系(RIL)群体71个株系和相应的全基因组染色体片段置换系(Chromosomesegmentsubstitutionline,CSSL)群体65个株系,在2种环境下对分蘖角度性状进行了数量性状基因座(QTL)定位和上位性效应的遗传分析。在两种群体中都出现了分蘖角度的超亲分离。在RIL群体中发现了5个主效QTLs和3对上位性双位点互作标记基因座,控制水稻分蘖角度。其中在第9染色体上位于XNpb108~C506RFLP分子标记区间的qTA-9基因座在2种环境中同时出现,其贡献率平均为28·6%,增加分蘖角度的等位基因来自籼稻品种IR24。利用CSSL群体图示基因型分析,证实在第9染色体上含有RFLP标记C609和C506约15cM的染色体区段,存在增加分蘖角度的基因,来源于染色体片段供体亲本IR24,在Asominori的遗传背景中能增加分蘖角度约15°,该基因的位置与RIL群体在第9染色体上定位的QTL相同,证实了qTA-9的存在。F1表型测定及F2代遗传分析表明,来自IR24的等位基因是一个不完全显性基因。除一对上位性位点存在显著的环境互作效应外,未发现其他位点存在与环境的互作效应。不同基因的加性效应和双位点的上位性效应的共同作用可能是造成水稻分蘖角度超亲分离的主要原因。     

水稻生物学产量及其构成性状的QTL定位

QTL的加性效应、加性×加性上位性效应及它们与环境的互作效应是数量性状的重要遗传分量.利用IR64/Azucena的125个DH品系为群体,分析了水稻生物学产量及其两个构成性状干草产量和谷粒产量的遗传组成.用基于混合模型的复合区间作图(MCIM)方法进行QTL定位.检测到12个位点有加性主效应,27个位点涉及双位点互作,18个位点存在环境互作.结果表明水稻生物学产量和它的两个构成性状普遍存在上位性效应和QE互作效应.此外,还探讨了性状间相关的遗传基础.发现4个QTLs和一对上位性QTLs可能与生物学产量与干草产量之间的正相关有关.3个QTL可能与干草产量与谷粒产量之间的负相关有关.这些结果可能部分地解释了这3个性状相关的遗传原因.通过对水稻生物学产量及其两个构成性状所定位QTL的分析,加深了对数量性状QTL的认识.首先,QTL的上位性效应和QE互作效应是普遍存在的;其次,QTL的多效性或紧密连锁可能是遗传相关的原因,当QTL对两个性状作用的方向相同时可导致正向遗传相关,反之则为负向遗传相关,当有些QTL表现为同向作用而另一些QTL表现为反向作用时,则可削弱性状间的遗传相关性;第三,复合性状的QTL效应可分解为其组成性状的QTL效应,如果QTL对各组成性状的效应方向相反而相互抵消,可使复合性状的QTL效应不易被检测;第四,加性效应的QTL常参预构成上位性效应,而具有上位性效应的QTL并非都有加性主效应,表明忽略上位性的QTL定位方法会降低检测QTL的功效;最后,鉴别不同类型的QTL效应有利于指导育种实践,选择主效QTL适用于多环境,QE互作QTL适用于特定环境,对上位性QTL应强调选择基因组合而并非单个基因.     

水稻单株有效穗数和每穗粒数的QTL剖析

应用292个Lemont/特青F13重组自交系（RILs)及其含272标记的遗传连锁图谱部析单株有效穗数（PN）和每穗粒数（SNP）及其相关性状的遗传,所有性状呈现超亲分离,PN与SNP存在弱的负相关,检测到影响PN和SNP及其相关性状的QTL51个和互作位点45对,它们可以解释60％以上的性状总变异,SNP与其相关性状的QTL定位在一起,比较与SNP－QTL同一或相邻区域的QTL数,穗部枝梗数是长度性状的两倍,故前者对SNP的作用更大,仅有2个PN－QTL与SNP相关性状的QTL相邻,因此通过标记辅助选择有可能实现PN与SNP的有利重组,其中影响PN的QPn4和影响穗枝梗数和长度的QPbn3a,QPbn3和QPb14等主效QTL,在标记辅助选择中具有重要的应用价值,对通过标记辅助选择合理配置穗部性状TQL产生新的高产穗型进行了讨论。     

Molecular mapping of genes conferring aluminum tolerance in rice (Oryza sativa L.)

Crop productivity on acid soil is restricted by multiple abiotic stress factors. Aluminum (Al) tolerance seems to be a key to productivity on soil with a pH below 5.0, but other factors such as Mn toxicity and the deficiency of P, Ca and Mg also play a role. The development of Al-tolerant genotypes of rice is an urgent necessity for improving crop productivity in developing countries. Inhibition of root growth is a primary and early symptom of Al toxicity. The present study was conducted to identify genetic factors controlling the aluminum tolerance of rice. Several parameters related to Al tolerance, most importantly the relative root growth under Al stress versus non-stress conditions, were scored in 188 F₃ selfed families from a cross between an Al-tolerant Vietnamese local variety, Chiembau, and an Al-susceptible improved variety, Omon269–65. The two varieties are both Oryza sativa ssp. indica, but showed a relatively high level of DNA polymorphism, permitting the assembly of an RFLP map consisting of 164 loci spanning 1,715.8 cM, and covering most of the rice genome. A total of nine different genomic regions on eight chromosomes have been implicated in the genetic control of root and shoot growth under aluminum stress. By far the greatest effects on aluminum tolerance were associated with the region near WG110 on chromosome 1. This region does not seem to correspond to most of the genes that have been mapped for aluminum tolerance in other species, nor do they correspond closely to one another. Most results, both from physiological studies and from molecular mapping studies, tend to suggest that aluminum tolerance is a complex multi-genic trait. The identification of DNA markers (such as WG110) that are diagnostic for aluminum tolerance in particular gene pools provides an important starting point for transferring and pyramiding genes that may contribute to the sustainable improvement of crop productivity in aluminum-rich soils. The isolation of genes responsible for aluminum tolerance is likely to be necessary to gain a comprehensive understanding of this complex trait. Received: 29 March 2000 / Accepted: 16 August 2000     

Development and mapping of 2240 new SSR markers for rice (Oryza sativa L.).

A total of 2414 new di-, tri- and tetra-nucleotide non-redundant SSR primer pairs, representing 2240 unique marker loci, have been developed and experimentally validated for rice (Oryza sativa L.). Duplicate primer pairs are reported for 7% (174) of the loci. The majority (92%) of primer pairs were developed in regions flanking perfect repeats > or = 24 bp in length. Using electronic PCR (e-PCR) to align primer pairs against 3284 publicly sequenced rice BAC and PAC clones (representing about 83% of the total rice genome), 65% of the SSR markers hit a BAC or PAC clone containing at least one genetically mapped marker and could be mapped by proxy. Additional information based on genetic mapping and "nearest marker" information provided the basis for locating a total of 1825 (81%) of the newly designed markers along rice chromosomes. Fifty-six SSR markers (2.8%) hit BAC clones on two or more different chromosomes and appeared to be multiple copy. The largest proportion of SSRs in this data set correspond to poly(GA) motifs (36%), followed by poly(AT) (15%) and poly(CCG) (8%) motifs. AT-rich microsatellites had the longest average repeat tracts, while GC-rich motifs were the shortest. In combination with the pool of 500 previously mapped SSR markers, this release makes available a total of 2740 experimentally confirmed SSR markers for rice, or approximately one SSR every 157 kb.     

QTL mapping for the paste viscosity characteristics in rice (Oryza sativa L.)

In order to understand the genetic basis of the paste viscosity characteristics (RVA profile, which is tested on the Rapid Visco Analyser) of the rice grain, we mapped QTLs for RVA profile parameters using a DH population derived from a cross between an indica variety, Zai-Ye-Qing 8 (ZYQ8), and a japonica variety, Jing-Xi 17 (JX17). Evidence of genotype-by-environment interaction was found by comparing the mapped QTLs between two locations, Hainan (HN) and Hangzhou (HZ). A total of 20 QTLs for six parameters of the RVA profiles were identified at least one location. Only the waxy locus (wx) located on chromosome 6 was detected significantly at both environments for five traits, i.e. hot paste viscosity (HPV), cool paste viscosity (CPV), breakdown viscosity (BDV), consistency viscosity (CSV) and setback viscosity (SBV). This locus explained 19.5%–63.7% of the total variations at both environments, suggesting that the RVA profiles were mainly controlled by the wx gene. HPV, CPV, BDV, CSV and SBV were also controlled by other QTLs whose effects on the respective parameter were detected only in one environment, while for the peak viscosity (PKV), only 2 QTLs, 1 at HN,the other at HZ, were identified. These results indicate that RVA profiles are obviously affected by environment. Received: 18 July 1999 / Accepted: 27 August 1999     

水稻对叶瘟和穗瘟部分抗性的遗传分析

在一个水稻籼籼交重组自交系群体中,选用由感病株系构成的2个亚群体和2个不同的稻瘟病菌小种,进行了水稻对叶瘟部分抗性的QTL定位,还选用由感病而且抽穗期相近的株系构成的亚群体和另一个病菌小种,进行了水稻对穗瘟部分抗性的QTL定位,将病叶面积百分比(DLA)、病斑大小(LS)和病斑数(LN)作为对叶瘟部分抗性的性状,将病斑长度(LL)和孢子量(CA)作为对穗瘟部分抗性的性状。所构建的图谱包含168个标记。应用QTLMapper 1．01b,共检测到11个表现主效应的QTL和28对双因子互作,有3个表现主效应的QTL参与对同一性状的互作。QTL的主效应对单一性状的贡献率为4．7％～38．8％,而上位性效应对单一性状的贡献率为16．0％～51．7％,QTL的主效应对大多数性状的贡献率小于互作效应,表明互作效应对于部分抗性的重要作用。对穗瘟部分抗性的两个性状LL和CA,所检测到QTL总效应的贡献率分别达到70．6％和82．6％,表明由排除了主效抗病基因的感病株系组成的亚群体适合于进行部分抗性QTL定位。     

栽培稻旱胁迫叶片相关性状的遗传解析

利用籼稻窄叶青8号(ZYQ8)和粳稻京系17(JX17)衍生的加倍单倍体(DH)群体127个株系,2002年在杭州采用田间断水法栽培,在水分胁迫下,对叶片的卷叶、相对含水量和电导率3个性状进行了评价和QTL分析。结果表明,3个性状在DH群体中均存在双向超亲分离,接近正态分布,受数量性状基因的控制;检测到影响这些性状的6个QTL,其中卷叶3个(qLR—1,qLR—5和qLR—11)、相对含水量2个(qRWC—1和qRWC—6和电导率1个(qERC—6）。旱胁迫时,目测卷叶方便易行,适于对大批品种或资源筛选,对抗旱栽培稻品种的筛选和利用具有一定的指导意义。     

水稻株高构成因素的QTL剖析

利用水稻籼粳杂交 (圭 6 30× 0 2 42 8) F1 的花药离体培养建立的一个含 81个 DH家系的作图群体 ,对水稻株高构成因素 (穗长、第 1节间长、……、第 5节间长 )进行基因定位。DH群体中株高构成因素均呈正态分布。相邻的构成因素间呈极显著的正相关 ,而相距较远的构成因素间的相关较弱或不显著。采用 QTL(Quantitative trait lo-cus)分析 ,定位了影响株高构成因素的 6个 QTL:qtl7同时影响穗长和第 1、2、3节间长 ,qtl1 和 qtl2 同时影响第 4和第 5节间长 ,qtl1 0 a和 qtl1 0 b仅影响第 1节间长 ,qtl3 仅影响第 3节间长。采用 QTL 互作分析 ,检测到 19对显著的互作 ,每个构成因素受 2个或 2个以上的 QTL 互作对的影响。并且还发现 ,同一个 QTL 互作对可能影响不同的性状 ,以及一个 QTL 可以分别与不同的 QTL 产生互作而影响同一个性状或影响不同的性状 ,但总的看来 ,加性效应是主要的。这些结果揭示了株高构成因素间相关的遗传基础 ,在水稻育种中运用这些 QTL 将有助于对株高 ,以及对穗长和上部节间长度进行精细的遗传调控。