Genotypic variation for tolerance to phosphorus deficiency in rice and the potential for its exploitation in rice improvement

Deficiency in phosphorus (P) can severely limit rice yields. Developing cultivars with tolerance to P deficiency may represent a more sustainable solution than sole reliance on fertilizer application. To assess genotypic variation for tolerance to P deficiency the P uptake of 30 genotypes was measured on P‐deficient soil. Variation for P uptake was high, ranging from 0.6 to 12.9 mg P/plant. Traditional varieties were superior to modern varieties. A major quantitative trait locus for P uptake had previously been identified in a population developed by crossing the modern variety ‘Nipponbare’ with the P deficiency‐tolerant landrace ‘Kasalath’. This quantitative trait locus was transferred to ‘Nipponbare’ by three backcrosses. Under P deficiency this improved line surpassed ‘Nipponbare’ in P uptake by 170% and in grain yield by 250%. These results show that the genotypic variation for tolerance to P deficiency in rice can be used successfully in rice improvement. By combining high P uptake of the donor variety ‘Kasalath’ with a high harvest‐index characteristic of modern varieties it was possible to more than triple the grain yield of ‘Nipponbare’ under P deficiency.     

Mapping of Re,a gene conferring the red leaf trait in ornamental kale (Brassica oleracea L. var. acephala)

Variegated leaf colour is an important agronomic trait that affects the market value of ornamental kale (Brassica oleracea L. var. acephala). The red leaf phenotype in kale is due to anthocyanin accumulation. To investigate the pattern of inheritance of this trait, we constructed an F₂ population by crossing ‘Y005‐15’, a double haploid with red leaves, with a white‐leaved double haploid, ‘Y011‐13‐38’, followed by self‐pollination. An F₂ population consisting of 4284 individuals was used to study the inheritance of this trait, which showed that the character was controlled by a dominate gene. All of the 1050 white leaf trait plants in the F₂ were used for mapping and developing markers linked to Re gene. Results showed that Re was mapped to a locus on linkage group C09 of Brassica oleracea, and the locus was mapped between six SSR markers (C9Z1, C9Z16‐1, C9Z90, C9Z94, C9Z96 and C9Z99), with a genetic distance of 6.7, 1.0, 0.3, 2.0, 2.1 and 0.4 cM from Re gene, respectively. These results may facilitate marker‐assisted selection of the red leaf trait in kale breeding as well as map‐based cloning of the red leaf trait gene.     

Identification of a locus for asynchronous heading in rice, <Emphasis Type="Italic">Oryza sativa</Emphasis> L.

We have identified a locus for asynchronous heading in rice. The length of heading period in a plant was measured as the indicator of asynchronous heading in 98 backcross inbred lines (BILs). These lines were derived using a single-seed descent method from a backcross of (japonica Nipponbare/indica Kasalath)/Nipponbare, and we used this mapping population in quantitative trait locus (QTL) analysis. We located a single QTL related to asynchronous heading (qah7) in the region of R2401–C39 on chromosome 7, and its contribution to the total variation was 0.14 (r²), with the positive effect due to the Kasalath allele. A near isogenic line (NILah7) that carries a Kasalath chromosomal segment corresponding to qah7 in the Nipponbare genetic background was selected and analyzed to identify a locus for asynchronous heading. The length of heading period in NILah7 was significantly longer (P < 0.001) than that in Nipponbare.     

QTL analysis of seed storability in rice

A double haploid population, which consists of 127 lines derived from anther culture of a typical indica and japonica hybrid ‘ZYQ8’/‘JX17’, was used in this study. Seed storability was investigated by using the storage property measured by the difference of seed germination rates before and after treatment of the rice seeds under 40°C and 95% relative humidity for 10 days in a phytotron. Three QTLs related to rice seed storability were detected on chromosomes 9, 11 and 12, with the LOD scores 2.76, 4.83 and 2.54, respectively, together explaining 35.4% of the genetic variation. The ‘JX17’ allele at qLS‐9 and the ‘ZYQ8’ alleles at qLS‐11 and qLS‐12 could enhance the rice seed storability. The effects of the ‘ZYQ8’ alleles of qLS‐11 and qLS‐12 were also verified using chromosome segment substitution lines.     

Fine mapping the QTL qFS‐chr.23, using a CSIL

In an earlier advanced‐backcross quantitative trait locus (QTL) analysis of an interspecific cross of Gossypium hirsutum cv. ‘Xinluzhong 36’(‘XLZH36’) and G. barbadense cv. ‘Xinhai 21’(‘XH21’), a QTL for fibre strength in the chromosome segment introgression line IL23‐09 was analysed. Single marker analysis revealed that the markers on chro.23 were associated with fibre strength. Using composite interval mapping with the F₂ population (1296 plants), a QTL for fibre strength was detected on chro. 23. The QTL explained 8.9% and 15.9% of phenotypic variances in the F₂ and F_2 : 3 generations, respectively. Substitution mapping suggested that the QTL was located at a physical distance of 23.4 kb between the markers BNL1414 and the single nucleotide polymorphism (SNP) locus D09_43776813 C‐G. We designated this QTL as qFS‐chr.23 (quantitative trait locus for fibre strength on chro.23). This work provides a valuable genetic resource for the breeding of high fibre quality in cotton and will facilitate future efforts for map‐based cloning.     

Deployment of cold tolerance loci from Oryza sativa ssp. Japonica cv. ‘Nipponbare’ in a high‐yielding Indica rice cultivar ‘93‐11’

Cold tolerance is a complex trait, and QTL pyramiding is required for rice breeding. In this study, a total of seven QTLs for cold tolerance in the Japonica rice variety ‘Nipponbare’ were identified in an F_2:3 population. A stably inherited major QTL, called qCTS11, was detected in the region adjacent to the centromere of chromosome 11. In a near‐isogenic line population, the QTL was further dissected into two linked loci, qCTS11.1 and qCTS11.2. Both of the homozygous alleles of qCTS11.1 and qCTS11.2 from ‘Nipponbare’ showed major positive effects on cold tolerance. Through pyramiding the linked QTLs in the cold‐sensitive Indica rice cultivar ‘93‐11’, we have developed a new elite, high‐yielding Indica variety with cold tolerance.     

Molecular mapping of a quantitative trait locus qSTV11Z harbouring rice stripe virus resistance gene,Stv‐b

Rice stripe virus (RSV) predominantly affects rice. In this study, we attempted to localize the quantitative trait locus (QTL) conferring RSV resistance in the ‘Zenith’ variety, which is known to harbour Stv‐a and Stv‐b. The resistant variety Zenith was crossed with the susceptible variety ‘Ilpum’ to generate a mapping population comprising 180 F_2:3 lines for QTL analysis. Contrary to previous findings, we could not detect Stv‐a‐specific QTLs on chromosome 6. Stv‐b‐specific QTL was detected on the long arm of chromosome 11; it was designated qSTV11^z. Six F_4:5 lines were selected from the F_3:4 population and fine‐mapped using insertion/deletion (InDel) markers. qSTV11^z was mapped to a 520‐kb region between the InDel markers Sid2 and Indel8. This region included OsSOT1 (candidate gene for STV11) and other previously reported RSV resistance QTLs. The OsSOT1 sequence in Ilpum and Zenith was identical to that of the susceptible variety ‘Koshihikari’, indicating that OsSOT1 is not the candidate gene of qSTV11^z. The localization of qSTV11^z should provide useful information for marker‐assisted selection and determination of genetic resources in rice breeding.     

控制水稻品种Koshihikari抽穗期的数量性状位点

利用Koshihikari × Kasalath BILs群体及相应的Kasalath/Koshihikari CSSLs群体,在江苏南京和海南陵水两个环境下对抽穗期的QTL定位分析。结果表明,在两地重复检测到3个QTL,分别位于第3、6和8染色体上;在qHd-3和qHd-8位点,来自Koshihikari等位基因能够提早抽穗,在qHd-6位点,来自Koshihikari等位基因能够延迟抽穗;第3染色体qHd-3和第7染色体非加性QTL之间存在上位性互作,通过重组自交系和置换系相互验证发现,qHd-3 所在的标记区间与W008的Kasalath插入片段位置大体一致,qHd-8所在的标记区间与W023、W024的Kasalath插入片段相吻合,qHd-7-1所在的标记区间位于W20的Kasalath片段之内,表明确实存在这3个位点。同时还对Koshihikari × 桂朝2号RILs抽穗期的QTL定位分析,在南京和海南分别检测到3个加性抽穗期QTL,1对非加性抽穗期QTL存在互作。     

Quantitative trait loci for leafhopper (Empoasca fabae and Empoasca kraemeri) resistance and seed weight in the common bean

A population of 108 common bean recombinant inbred lines (RILs) (F_5:6‐9), derived from a leafhopper (Empoasca fabae and E. kraemeri)‐susceptible cultivar (‘Berna’) and a leafhopper‐resistant line (EMP 419) was used to identify molecular markers genetically linked to leafhopper resistance and seed weight. Bulked segregant analysis and quantitative trait analysis identified eight markers that were associated with resistance to E. fabae, and four markers that were associated with E. kraemeri resistance. Three markers were associated with resistance to both species. A partial linkage map of the bean genome was constructed. Composite interval mapping identified quantitative trait loci (QTL) for resistance to both leaf hopper species on core‐map linkage groups B1, B3 and B7. QTL for seed weight were found close to the locus controlling testa colour and an α‐phaseolin gene.     

Identification of a major blast resistance gene in the rice cultivar 'Tetep'

Analysis of near‐isogenic lines (NILs) indicated the presence of a novel resistance gene in the indica rice cultivar ‘Tetep’ which was highly resistant to the rice blast fungus Magnaporthe grisea.‘Tetep’ was crossed to the widely used susceptible cultivar ‘CO39’ to generate the mapping population. A Mendelian segregation ratio of 3 : 1 for resistant to susceptible F₂ plants further confirmed the presence of a major dominant locus, in ‘Tetep’, conferring resistance to the blast fungal isolate B157, corresponding to the international race IC9. Simple sequence length polymorphism (SSLP) was used for molecular genetic analysis. The analysis revealed that the SSLP marker RM 246 was linked to a novel blast resistance gene designated Pi‐tp(t) in ‘Tetep’.     

Identification and validation of an over‐dominant QTL controlling soybean seed weight using populations derived from Glycine max × Glycine soja

Heterosis, or hybrid vigour, has been used to improve seed yield in several important crops for decades and it has potential applications in soybean. The discovery of over‐dominant quantitative trait loci (QTL) underlying yield‐related traits, such as seed weight, will facilitate hybrid soybean breeding via marker‐assisted selection. In this study, F₂ and F_2 : 3 populations derived from the crosses of ‘Jidou 12’ (Glycine max) × ‘ZYD2738’ (Glycine soja) and ‘Jidou 9’ (G. max) × ‘ZYD2738’ were used to identify over‐dominant QTL associated with seed weight. A total of seven QTL were identified. Among them, qSWT_13_1, mapped on chromosome 13 and linked with Satt114, showed an over‐dominant effect in two populations for two successive generations. This over‐dominant effect was further examined by six subpopulations derived from ‘Jidou12’ × ‘ZYD2738’. The seed weight for heterozygous individuals was 1.1‐ to 1.6‐fold higher than that of homozygous individuals among the six validation populations examined in different locations and years. Therefore, qSWT_13_1 may be a useful locus to improve the yield of hybrid soybean and to understand the molecular mechanism of heterosis in soybean.     

QTL mapping and associated marker selection for the efficacy of green plant regeneration in anther culture of rice

Anther culturability of rice is a quantitative trait controlled by nuclear‐encoded genes. The identification of quantitative trait loci (QTL) and associated marker selection for anther culturability is important for increasing the efficiency of green plant regeneration from microspores. QTL associated with the capacity for green plant regeneration in anther culture of rice were mapped on chromosomes 3 and 10 using 164 recombinant inbred (RI) lines from a cross between ‘Milyang 23’ and ‘Gihobyeo’. The quantitative trait locus located on chromosome 10 was detected repeatedly when three anther culture methods were applied and was tightly linked to the markers, RG323, RG241 and RZ400. Associations between these markers and the efficacy of green plant regeneration in 43 rice cultivars and two F₂ populations, ‘MG RI036’/‘Milyang 23’, and ‘MG RI036’;/‘IR 36’ were analysed. One of these markers, RZ400, was able to identify effectively genotypes with good (> 10.0%) and poor (< 3.0%) regenerability, based on the marker genotypes in the cultivars and two F₂ populations. This marker enables the screening of rice germplasm for anther culturability and introgression into elite lines in breeding programmes.     

Identification of quantitative trait loci for resistance to bending-type lodging in rice (Oryza sativa L.)

Bending-type lodging is one of the most important factors affecting the yield and grain quality of rice. This study identified quantitative trait loci (QTLs) for physical strength of the upper culms, and evaluated QTL effects on lodging resistance. In 2010 and 2011, QTLs for breaking strength, length, and diameter of the top three internodes were identified by analyzing chromosomal segment substitution lines (CSSLs) developed from ‘Koshihikari’ and ‘Kasalath’. The QTL analysis indicated that ‘Kasalath’ had two types of QTLs: one to strengthen specific internodes and one to simultaneously improve the physical strengths of plural internodes or the top three internodes. A QTL for breaking strengths of the top three internodes (bsuc11) was detected on chromosome 11 in both years. This QTL did not overlap with that for internode length. To evaluate the effects of bsuc11 on lodging resistance, this study selected three CSSLs with bsuc11 and analyzed the breaking strengths of the top three internodes after heading and the pushing resistance of the lower part. Internodes of ‘Koshihikari’ showed decreased breaking strengths after grain filling, while those of CSSLs with bsuc11 did not show this decrease in breaking strength. The pushing resistance of the lower part at the fully ripe stage was the same in ‘Koshihikari’ and CSSLs with bsuc11. These results suggested that bsuc11 could be a target to improve the physical strength of the upper culms to resist bending-type lodging, and that the physical strengths of upper and lower parts are controlled by different genetic factors in rice.     

Development of cleaved amplified polymorphic sequence (CAPS) markers linked to a green rice leafhopper resistance gene, Grh3(t)

The chromosomal location of the resistance gene for green rice leafhopper (GRLH), an injurious insect for rice, has been determined and RFLP markers closely linked to this gene have been identified. The susceptible japonica rice variety ‘Nipponbare’ was crossed with a resistant japonica rice line ‘Aichi42’, in which green rice leaf hopper resistance had been introduced from an indica variety ‘Rantaj‐emas2’, and the 100 F₂ plants obtained were used for linkage analysis. The green rice leafhopper resistance gene, Grh3(t), was mapped between RFLP markers C288B and C133A on chromosome 6 and co‐segregated with C81. Of the RFLP markers tightly linked to Grh3(t), C81 was converted to a SCAR marker and C133A to a cleaved amplified polymorphic sequence marker that could distinguish the heterozygous genotype to establish an effective marker‐aided selection system for the GRLH resistance gene.     

A novel aphid resistance locus in cowpea identified by combining SSR and SNP markers

The utility of combining simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) marker genotyping was determined for genetically mapping a novel aphid (Aphis craccivora) resistance locus in cowpea breeding line SARC 1‐57‐2 and for introgressing the resistance into elite cultivars by marker‐assisted backcrossing (MABC). The locus was tagged with codominant SSR marker CP 171F/172R with a recombination fraction of 5.91% in an F₂ population from ‘Apagbaala’ x SARC 1‐57‐2. A SNP‐genotyped biparental recombinant inbred line population was genotyped for CP 171F/172R, which was mapped to position 11.5 cM on linkage group (LG) 10 (physical position 30.514 Mb on chromosome Vu10). Using CP 171F/172R for foreground selection and a KASP‐SNP‐based marker panel for background selection in MABC, the resistance from SARC 1‐57‐2 was introduced into elite susceptible cultivar ‘Zaayura’. Five BC₄F₃ lines of improved ‘Zaayura’ that were isogenic except for the resistance locus region had phenotypes similar to SARC 1‐57‐2. This study identified a novel aphid resistance locus and demonstrated the effectiveness of integrating SSR and SNP markers for trait mapping and marker‐assisted breeding.     

Mapping of quantitative trait loci controlling seedling vigor in rice (Oryza sativa L.) under submergence

Submergence-induced suppression of seedling vigor is a serious constraint particularly in the direct seeding rice cultivation system. To identify quantitative trait loci (QTLs) associated with seedling vigor in rice under submergence, a mapping population of 98 Backcross Inbred Lines derived from a cross of Nipponbare/Kasalath//Nipponbare was used. Phenotypic evaluation of seedling vigor under submergence was based on shoot length (SHL), root length (RTL) and shoot fresh weight (SFW) using a test tube bioassay method. Thirty-two putative QTLs were detected among which 7 were for SHL, 11 for RTL and 14 for SFW. Phenotypic evaluation was also made of the parental lines and a set of 54 chromosome segment substitution lines in which Nipponbare segments were substituted for by their homologous Kasalath segments covering the entire rice genome. Two QTLs with more than 10 % contribution to the total phenotypic variance were verified for SHL, and at least one for RTL and six for SFW on chromosomes 1, 3, 4, 6 and 7 at the 1 % significance level. Among these, all but two showed reductions in one, two or all three traits. Our present and previous results suggest that the Nipponbare genome has a potential to improve seedling vigor under submergence and that japonica germplasms can be used to breed for this important trait in indica rice.     

A new locus for resistance to brown planthopper identified in the indica rice variety DV85

The brown planthopper (BPH) is one of the most destructive insect pests of rice. Resistant varieties have proved to be one of the most economic and effective measures for BPH management. In this study, an indica rice ‘DV85’ showed resistance to biotype 2 of BPH by bulked seedling test, and a recombinant inbred line (RIL) population derived from a cross between a susceptible rice ‘Kinmaze’ and ‘DV85’ was phenotyped to map genetic factors conferring BPH resistance in ‘DV85′. Composite interval mapping revealed that one quantitative trait locus (QTL) with a LOD score of 10.1 was detected between XNpb202 and C1172 on chromosome 11. This QTL was designated as Qbph11. Qbph11 explained 68.4% of the phenotypic variance of BPH resistance in this population. The allele from the resistant parent ‘DV85’ at Qbph11 reduced the damage caused by BPH feeding and would be very useful in breeding resistant rice varieties via marker‐assisted selection.     

Dissection of genetic variance of fibre quality in advanced generations from an interspecific cross of Gossypium hirsutum and G. barbadense

The interspecific genetic introgression approach has been shown to facilitate the detection and dissection of quantitative trait loci (QTL). A population consisting of 121 F₆ recombinant inbred lines was developed by crossing Gossypium hirsutum cv. ‘Handan 208’ and G. barbadense cv. ‘Pima 90’ via single‐seed descent. Genotyping indicated that the mean ‘Pima 90’ allele frequency at each locus was 21%. Phenotyping and phenotypic distribution indicated a trend of return of individual lines’ characters to ‘Handan 208’ coupled with a wide variance for each trait. Significant loci influencing fibre quality were detected by one‐way analysis of variance (anova; P < 0.005) and association analysis [?log₁₀(P) ≥ 3]: five and three markers for fibre length, four and one marker(s) for uniformity, two and one marker(s) for micronaire, 13 and 15 markers for strength, six and 10 markers for elongation, respectively. Two‐way anova based on genotypes of all marker loci combination showed that 807 two‐locus combinations were significant, and two‐way anova based on marker genotypes of QTL markers combination showed five significant digenic interactions (P < 0.01).     

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.