Mapping of QTLs for leaf developmental behavior in rice (Oryza sativa L.)

Leaf developmental behavior in rice (Oryza sativa L.) is one of the important agronomic characteristics, which not only determines vegetative growth but also influences grain yield. This study was conducted to identify quantitative trait loci (QTLs) for total number of leaves (TNL), days to the emergence of flag-leaf (DEF) and the leaf emergence rates (LER) on main stem, which mainly represent leaf developmental behavior, using recombinant inbred lines (RILs) derived from a cross between a japonica variety, Asominori and an indica variety, IR24, cultivated in 2001 and 2002. The transgressive segregations in both parental directions and continuous variations of all three tested traits were observed. Significant correlations among these traits were detected. A total of fourteen QTLs for leaf development behavior were detected with 289 RFLP markers. Six QTLs controlling TNL were mapped to chromosomes 3, 5, 6, 8, 9, 12, and accounted for 5.615.7% of the total phenotypic variations, and three QTLs for DEF were mapped to chromosome 3, 6, 8 and accounted for 10.735.4% of total phenotype variation and five QTLs for LER were mapped to chromosome 1(two QTLs), 2, 4, 9 and explained 6.217.5% of phenotype variation. The identification of QTLs for leaf developmental behavior in rice may be useful for selection of fast growing genotype before heading using maker-assisted selection.     

Association analysis of physicochemical traits on eating quality in rice (Oryza sativa L.)

Improvement of rice eating quality is an important objective in current breeding programs. In this study, 130 rice accessions of diverse origin were genotyped using 170 SSR markers to identify marker–trait associations with physicochemical traits on eating quality. Analysis of population structure revealed four subgroups in the population. Linkage disequilibrium (LD) patterns and distributions are of fundamental importance for genome-wide mapping associations. The mean r ² value for all intrachromosomal loci pairs was 0.0940. LD between linked markers decreased with distance. Marker–trait associations were investigated using the unified mixed-model approach, considering both population structure (Q) and kinship (K). In total, 101 marker–trait associations (p < 0.05) were identified using 52 different SSR markers covering 12 chromosomes. The results suggest that association mapping in rice is a viable alternative to quantitative trait loci mapping, and detection of new marker–trait associations associated with rice eating quality will also provide important information for marker-assisted breeding and functional analysis of rice grain quality.     

利用东乡普通野生稻染色体片段置换系定位产量相关性状QTL

以东乡普通野生稻和日本晴为亲本构建的染色体片段置换系为研究材料, 2019年分别在北京、山东临沂和江西南昌对分蘖数、穗粒数和粒形等11个产量相关性状进行多环境鉴定,结合染色体片段置换系基因型数据定位水稻产量相关性状QTL。3个环境共检测到68个QTL,包括株高4个、穗长5个、分蘖数2个、一次枝梗数7个、一次枝梗粒数8个、二次枝梗数8个、二次枝梗粒数10个、每穗粒数6个、千粒重7个、粒长8个和粒宽3个; LOD值介于2.50～12.66之间,贡献率变幅为4.67%～27.79%,15个QTL的贡献率大于15%;24个QTL与已报道位点/基因位置重叠,44个QTL为新发现位点; 6个QTL在2个环境能被检测到, 1个QTL qTGW2能在3个环境检测到,且是还未报道的新位点。最后,利用BSA法验证了qPH7、qPBPP8-2和qGW10三个QTL的可靠性。本研究将为后续产量相关性状基因克隆以及进一步解析其遗传基础和分子调控机制奠定基础。     

水稻幼苗活力相关性状的QTLs定位和上位性分析

以籼粳交IR64/Azucena的DH群体109个株系为研究材料, 在黑暗条件下, 通过适温(25～28℃)和低温逆境(16～19℃)对该DH群体的种子进行发芽试验, 测定幼苗中胚轴长度、芽鞘长、根长、苗高等幼苗活力的相关性状. 采用QTLMapper统计软件检测控制这些性状的加性效应QTLs和加性×加性上位性QTLs, 共检测到24个加性效应QTLs和17对上位     

基于CSSSLs的水稻穗长QTL的定位

穗长是影响水稻产量的重要因子之一,是典型的数量性状,遗传基础复杂,且易受环境等因素的影响。染色体单片段代换系(Chromosome single segment substitution lines,CSSSLs)减少了个体间遗传背景的干扰,是鉴定复杂性状QTL的新型遗传材料。本研究以广陆矮4号为受体、日本晴为供体的85个染色体单片段代换系群体为试验材料,通过单因素方差分析和Dunnett’s多重比较测验单片段代换系与受体亲本广陆矮4号之间穗长的差异,对代换片段上穗长QTL进行了鉴定。以P<0.001为阈值,共检测到22个穗长QTLs,分布于除第10染色体以外的11条染色体上,其加性效应值的变化范围为-2.63～3.87,加性效应百分率变化范围为-11.47%～16.88%。这些QTLs的鉴定,为进一步克隆穗长QTL以及水稻穗长的分子改良提供了重要的依据。     

Substitution mapping of QTLs for blast resistance with SSSLs in rice (Oryza sativa L.)

Rice blast, caused by fungal pathogen Magnaporthe grisea Barr., is one of the most devastating rice diseases worldwide. It has greatly affected the rice production and quality. Development of resistant cultivars is the most effective and economical way for controlling this disease in rice. In this study, 114 of the single-segment substitution lines (SSSLs) in rice were inoculated at seedling stage by 16 rice blast isolates. The substituted segments in the 114 SSSLs distributed on 12 chromosomes with coverage of 57.32% of rice genome. Fifteen of the SSSLs were different in blast resistance from the HJX74 recipient. The SSSL W23-7-6-5-2-2 showed 100% of resistance frequency. A total of 11 QTLs for blast resistance were detected on chromosomes 1, 2, 3, 6, 10, 11 and 12 in rice. They were mapped at chromosomal intervals of 2.2?C46.2?cM, of which 6 QTLs were mapped at less than 10.0?cM. Six of the 11 QTLs were first reported in this paper.     

稻米直链淀粉含量基因座位的分子标记定位

以直链淀粉含量(AC)中等的CT9993和泰国优质的香软米KDML105杂交产生的152个重组近交系 (RIL)为材料, 构建了含83个RFLP、 69个AFLP和15个微卫星(SSLP)标记的分子标记连锁图 , 标记间平均距离为12.98 cM。 应用该连锁图对控制稻米AC的基因座位(QTL)进行了分析 。 结果表明： 稻米AC主要受两个主效QTL和5个微效QTL的共同控制     

Detection and validation of QTLs associated with seed longevity in rice (Oryza sativa L.)

Seed longevity in rice is a major determinant in seed production and germplasm preservation. In this paper, a recombinant inbred line (RIL) population consisting of 172 lines derived from the cross between Xiang743 and ‘Katy’ was used to map quantitative trait loci (QTLs) for seed longevity (SL) after seed storage for 18 and 30 months under ambient conditions. Two putative QTLs, qSL‐2 and qSL‐8, were detected and located on chromosomes 2 and 8, respectively. qSL‐2 is an allele from Xiang743 allele and increases seed longevity. qSL‐8 was a novel QTL from ‘Katy’ allele and increases seed longevity. qSL‐8 explained 15.29% and 17.35% of the phenotypic variance after seed storage for 18 and 30 months, respectively. Furthermore, qSL‐8 was validated in a secondary population developed by self‐pollination of a residual heterozygous line (RHL) selected from the RIL population, which explained 25.93% of the phenotypic contribution. These results provide an opportunity for map‐based cloning of qSL‐8. Furthermore, qSL‐8 may be a target for improving seed longevity by marker‐assisted selection (MAS) in rice.     

水稻株高及其构成因素数量性状基因座位的分子标记定位

应用具有８９个标记位点的Ｆ２群体的ＲＦＬＰ图谱，对控制水稻株高及其构成因素的数量性状基因座位进行定位等研究。共定位了３２个ＱＴＬｓ，其中涉及株高的有７个、穗长３个、第一节间长有２个、第二节间长和第三节间长各３个、第四节间长有６个、节间数和第一节间干重各有４个。     

Linkage studies in rice (Oryza sativa L.)

Summary In an intervarietal cross between IR 8 and Wanar 1, inheritance of pigmentation in coleoptile, leaf-sheath, auricle, node, internode, septum and glume was studied. A pleiotropic gene conditioning pigment in all these parts has been identified which may probably be one of the basic genes for anthocyanin pigmentation. A linkage group consisting of genes causing pigment in these seven organs has been indicated in this sequence: Pau-Pc-Pn-Psh-Pin-Pg-Pm. This group may form part of the III Sp group of Misro et al. (1966) (corresponding to the A linkage group of japonica) in which case Pau, Pc, Pn and Pg are additions to this group. Linkage relationship of genes for coleoptile colour is reported for the first time.     

Wide compatibility in rice (Oryza sativa L.)

Summary Wide compatible varieties (WCVs) show normal spikelet fertility in crosses with Indica and Japonica rice varieties. Crosses of Indica and Japonica varieties frequently show high spikelet sterility which prevents exploitation of heterosis for grain yield. We screened 41 rice varieties for the wide compatibility trait by crossing each with three Indica and three Japonica testers. Varieties giving fertile F₁ hybrids with both groups of testers were classified as WCVs. Seven varieties viz., BPI-76 (Indica); N 22; Lambayeque-1 and Dular (Aus); Moroberekan, Palawan and Fossa HV (Japonicas), were identified as WCVs. The frequency of WCVs was higher among Aus and Japonicas. The wide compatibility trait in varieties: Dular and Moroberekan was controlled by a single dominant gene linked with the Est-2 and Amp-3 loci (mean recombination 32.0%). Est-2 and Amp-3 showed complete linkage. Pgi-2 was found to be linked with Est-2 and Amp-3 (mean recombination 16.1%). Est-2 and Amp-3, showed a tighter linkage with C ⁺ (mean recombination 4.1%). Pgi-2 showed a lower linkage with C ⁺ (mean recombination 17.3%). The recombination values between the WC gene in Dular and C ⁺ was much higher than those reported in Japan for the WC gene (S₅ ⁿ) from Ketan Nangka. It is possible that the WC gene from Dular is different from that in Ketan Nangka. Linkage intensities with the WC gene were not strong enough to be of use for indirect selection for the wide compatibility trait. A search for a more closely linked isozyme or DNA marker was proposed.     

Fine mapping of stable QTLs related to eating quality in rice (Oryza sativa L.) by CSSLs harboring small target chromosomal segments

Amylose content (AC) and viscosity profile are primary indices for evaluating eating and cooking qualities of rice grain. Using chromosome segment substitution lines (CSSLs), previous studies identified a QTL cluster of genes for rice eating and cooking quality in the interval R727–G1149 on chromosome 8. In this study we report two QTLs for viscosity parameters, respectively controlling setback viscosity (SBV) and consistency viscosity (CSV), located in the same interval using rapid viscosity analyzer (RVA) profile as an indicator of eating quality. Previously reported QTL for AC was dissected into two components with opposite genetic effects. Of four QTLs, qCSV-8 and qAC-8-2 had stable genetic effects across three and four environments, respectively. qSBV-8, qCSV-8 and qAC-8-1 partly overlapped, but were separated from qAC-8-2. Based on data from an Affymetrix rice GeneChip, two genes related to starch biosynthesis at the qAC-8-2 locus were chosen for further quantitative expression analysis. Both genes showed enhanced expression in sub-CSSLs carrying the target qAC-8-2 allele, but not in sub-CSSLs without the target qAC-8-2 allele, indicating their possible role in rice quality determination. Molecular markers closely linked to the two stable QTL provide the opportunity for marker-assisted selection (MAS) in breeding high quality rice.     

基于染色体单片段代换系的水稻粒形QTL定位

水稻的粒形是影响水稻产量和品质的重要因子之一, 是由多基因控制的数量性状。染色体单片段代换系由于减少了个体间遗传背景的干扰, 已经成为鉴定复杂性状QTL的新型遗传材料。本研究以广陆矮4号为受体,日本晴为供体的119个染色体单片段代换系群体为试验材料,通过单因素方差分析和Dunnett’s多重比较,测验单片段代换系与受体亲本之间粒形的差异,鉴定了代换片段上粒形相关的QTL。以P≤0.001为阈值, 共检测到39个粒形相关的QTL。其中,粒长相关的19个,其加性效应值为0.18~1.06 mm,加性效应百分率为2.40%~14.13%;粒宽相关的14个,其加性效应值为0.09~0.31 mm,加性效应百分率为2.71%~9.15%;粒厚相关的6个,其加性效应值为0.05~0.10 mm,加性效应百分率为2.14%~4.46%。这些QTL的鉴定,为进一步精细定位并克隆相应QTL和高产、优质水稻新品种的分子标记辅助选择奠定了基础。     

水稻单片段替换系群体的建立及QTL定位

水稻中许多重要的农艺性状属数量性状，由多基因(QTL)控制。研究QTL的遗传特性和遗传效应对于培育高产和稳产的水稻品种具有十分重要的意义。本研究以6个优良的品种为供体亲本，以华粳籼74为轮回亲本，通过微卫星标记辅助回交选择培育了一批单片段替换系，随后利用所培育的单片段替换系进行了QTL分析和基因定位。主要结果有：1、利用258个微卫星标记对6个供体亲本和轮回亲本间的多态性进行了筛选。6个供体亲本与轮回亲本间的多态率在32．98％至60．78％之间，平均47．81％，粳型供体亲本比籼型供体亲本的多态性要高。2、随着回交代数的增加，植株所含的替换片段数逐渐减少。在BC2F1、BC3F1、BC3F2和BC3F3代，平均每个植株携带有12．50、5．98、1．69和1．46个替换片段。替换片段的平均长度也随回交和自交代数的增加而逐渐变短。在BC2F1、BC3F1、BC3F2和BC3F3代，替换片段的平均长度分别为25．43cM、22．38cM、20．78cM和18．15cM.回交世代替换片段变短的速率(11．99％)比自交世代变短速率(7．15％)要快。在BC2F1、BC3Fl、BC3F2和BC3F3代，轮回亲本基因组的恢复率分别为82．24％、92．55％、98．04％j阳98．52％。3、在BC3F2和BC3F3代，共选育出111个单片段替换系，其中独一无二的单片段替换系共42个。BC3F2代替换系中替换片段的估算长度在2．00cM到64．80cM之间，平均为21．75cM，而BC3F3代中替换片段的估算长度在6．05cM到48．90cM之间，平均为20．95cMc，12条染色体中仅第11染色体没有选择到单片段替换系。所选育的单片段替换系中替换片段的总长为2367．50cM，基因组的覆盖长度为704．50cM，覆盖率为39．25％。4、在52个单片段替换系的22个性状中共鉴定出了234个QTL。每个性状鉴定出的QTL数在3到19个之间，平均4．50个。每个替换系鉴定出的QTL在2到15个之间，平均10．64个。QTL加性效应的大小因性状和替换系不同而不同，低至-0．02(0．79％)的加性效应(如谷粒宽度)均能检测到。在RM237．RM322、RM225和RM481标记附近的各种性状中同时检测到了增效和减效的QTL。5、通过染色体替换作图，对7个单片段替换系中16个性状的QTL进行了定位。利用携带有相同替换片段的替换系进行QTL位置的确定。6、通过分析复杂性状的加性效应，对影响植高、每穗粒数和粒型的相关性状进行了分析，分析QTL之间的相关性。7、对控制抽穗期、稃尖颜色、植株高度和粒型的基因进行了定位。抽穗期基因Hd-8表现为单基因显性遗传，定位于第8染色体上，与PSMl55紧密连锁。紫色稃尖基因Pa-6表现为单基因显性遗传，定位于第6染色体上，与RM253紧密连锁。株高基因Ph-1-3定位于第1染色体上，与PSM331紧密连锁。粒型基因Rlw-8-2为首次报道，定位于第8染色体的末端，与RM447紧密连锁，长粒表现为单基因隐性遗传。本研究通过分子标记辅助选择培育了一批单片段替换系，并成功地对这批材料进行了QTL分析和基因定位，从而证实了在水稻中通过微卫星标记辅助回交选育单片段替换系和进行QTL分析的可行性。     

Linkage studies in rice (Oryza sativa,L.)

Linkage studies in rice up to 1959 have been briefly reviewed with a special reference to the work carried out at the Central Rice Research Institute, Cuttack, India. The crossover values for several genes have been furnished. Instances have been cited wherein two or more genes appear to be involved in determining the expression of anthocyanin pigmentation in different plant parts.

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Samenvatting Een overzicht wordt gegeven van de literatuur tot en met 1959 over de koppeling van genen bij rijst, in het bijzonder het koppelingsonderzoek uitgevoerd door het Central Rice Research Institute, Cuttack, India.De crossover-waarden voor vele genen worden vermeld. Enkele gevallen zijn genoemd waarbij 2 of meer genen een pleiotrope werking hebben bij de vorming van anthocyaan in verschillende delen van de plant.

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Paper read in the meeting of the Working Party on Rice Production and Protection of the International Rice Commission, held in Ceylon during December, 1959.     

Effects of cytoplasm on heterosis and combining ability for agronomic traits in rice (Oryza sativa L.)

Summary Effect of sterilizing (WA) cytoplasm on heterosis and combining ability for days to flowering, plant height and grain yield in rice was studied in 70 crosses and their reciprocals produced by 10 cytoplasmic male sterile (A), their maintainer (B) and seven restorer (R) lines following line x tester design. The materials consisting of 140 hybrids (70 A/R and their reciprocal 70 R/B) plus 17 parental lines (10 B + 7 R) were evaluated under six environments, created by growing in three fields with different fertilizer doses (0, 60, 120 kg N/ha) and 2 seasons (dry and wet) during 1986 at IRRI farm. Reciprocal cross effect (A/R vs R/B) were highly significant for all the three traits. Interaction of reciprocal cross effects with environments were also highly significant for yield and days to flowering. Cytoplasmic effect for yield, days to flowering and plant height were estimated by comparing A/R and R/B combination and testing the significance of difference with LSD value. In order to avoid confounding effect of spikelet sterility on yields, twenty crosses showing normal spikelet fertility were selected. Both positive and negative cytoplasmic effects were observed for the three traits. The effects were modified by environments, except for plant height. Heterosis for all three traits was also affected by cytoplasm, however, manifestation of cytoplasmic effects was higher for heterosis for days to flower than in heterosis for yield and plant height. Effect of cytoplasm was more pronounced on general combining ability effects of parents than specific combining ability effects of crosses. Among the parents two CMS lines A4 (IR54752A) and A8 (IR22107-113-3-3A) and two R lines: R2 (IR46) and R7 (IR9761-19-1) showed consistent positive effect of cytoplasm on general combining ability. These lines have given several good heterotic combinations. The study indicated the usefulness of evaluating diverse cytoplasmic sources in various nuclear genotypes bred for hybrid rice breeding program.     

利用4个姊妹近等基因系群体定位水稻粒重和粒形QTL

粒重是决定水稻产量的三要素之一。利用世界上粒重最大的品种之一SLG-1(供体亲本)与小粒品种日本晴(Nipponbare,轮回亲本)杂交,在各回交世代选择粒重较大单株与日本晴回交,构建水稻粒重和粒形的姊妹近等基因系(SNILs)。对获得的73株BC4F1单株进行粒重频率分布统计,选择粒重频率分布在4个峰值处的代表性单株,自交获得4个BC4F2SNILs群体。利用BSA法(分离群体分组混合分析法),从均匀分布在水稻染色体上的1513对SSR标记中筛选出与粒重和粒形相关的多态性标记19对,以LOD≥2.5作为选择阈值,对粒重、粒长、粒宽和粒厚进行QTL扫描,共检测到6个区域的12个QTL,贡献率从7.22%到53.38%。这些QTL所在区域包含已克隆的粒长GS3和粒宽GW2,也包含没有精细定位的第2染色体的RM6318～RM1367、第3染色体的RM5477～RM6417和第6染色体的RM3370～RM1161等3个区域控制粒重和粒形的5个QTL。其中第3染色体上RM5477～RM6417区间存在粒形贡献率较大的新的QTL。构建含有这些粒重QTL的姊妹近等基因系,为进一步精细定位或克隆新的粒重或粒形QTL奠定了基础。