Analyses and identifications of quantitative trait loci and candidate genes controlling mesocotyl elongation in rice

Rice direct seeding has the significant potential to save labor and water,conserve environmental resources,and reduce greenhouse gas emissions tremendously.Therefore,rice direct seeding is becoming the major cultivation technology applied to rice production in many countries.Identifying and utilizing genes controlling mesocotyl elongation is an effective approach to accelerate breeding procedures and meet the requirements for direct-seeded rice(DSR) production.This study used a permanent mapping population with 144 recombinant inbred lines(RILs) and 2 828 bin-markers to detect quantitative trait loci(QTLs) associated with mesocotyl length in 2019 and 2020.The mesocotyl lengths of the rice RILs and their parents,Lijiangxintuanheigu(LTH) and Shennong 265(SN265),were measured in a growth chamber at 30°C in a dark environment.A total of 16 QTLs for mesocotyl length were identified on chromosomes 1(2),2(4),3(2),4,5,6,7,9,11(2),and 12.Seven of these QTLs,including qML1a,qML1b,qML2d,qML3a,qML3b,qML5,and qML11b,were reproducibly detected in both years via the interval mapping method.The major QTL,qML3a,was reidentified in two years via the composite interval mapping method.A total of 10 to 413 annotated genes for each QTL were identified in their smallest genetic intervals of 37.69 kb to 2.78 Mb,respectively.Thirteen predicted genes within a relatively small genetic interval(88.18 kb) of the major mesocotyl elongation QTL,qML3a,were more thoroughly analyzed.Finally,the coding DNA sequence variations among SN265,LTH,and Nipponbare indicated that the LOC_Os03g50550 gene was the strongest candidate gene for the qML3a QTL controlling the mesocotyl elongation.This LOC_Os03g50550 gene encodes a mitogen-activated protein kinase.Relative gene expression analysis using qRT-RCR further revealed that the expression levels of the LOC_Os03g50550 gene in the mesocotyl of LTH were significantly lower than in the mesocotyl of SN265.In conclusion,these results further strengthen our knowledge about rice’s genetic mechanisms of mesocotyl elongation.This investigation’s discoveries will help to accelerate breeding programs for new DSR variety development.

Identification of Quantitative Trait Loci for Anthesis-Silking Interval and Yield Components Under Drought Stress in Maize

A genetic linkage map with 89 SSR marker loci was constructed based on a maize (Zea mays L.) population consisting of 184 F-2 individuals from the cross, Huangzao 4 X Ye 107. The 184 F-3 families were evaluated in the field under well-watered and drought-stressed regimes in Shanxi Province of China. The objectives of the study were to identify genetic segments responsible for the expression of anthesis-silking interval (ASI), ear setting and grain yield, and to examine if the quantitative trait loci (QTLs) for ASI or yield components can be used in marker-assisted selection (MAS) to improve grain yield under drought conditions. Results showed that under well-watered and drought-stressed regimes, three and two QTLs involved in the expression of ASI were detected on chromosomes 1, 2 and 3, and 2 and 5, respectively. Under well-watered regime, two QTLs for ear setting were detected on chromosomes 3 and 6, explaining about 19.9% of the phenotypic variance, and displayed additive and partial dominant effects, respectively. Under drought-stressed condition, four QTLs for ear setting were detected on chromosomes 3, 7 and 10, which were responsible for interpreting 60.4% of the phenotypic variance, and showed dominant or partial dominant effects. Under well-watered condition, four QTLs controlling grain yield were identified on chromosomes 3, 6 and 7, while five QTLs were identified under drought stress on chromosomes 1, 2, 4 and 8. The gene action was of additive or partial dominant effects, and each QTL could explain 7.3% to 22.0% of the phenotypic variance, respectively. Under drought conditions, ASI and ear setting percentage were highly correlated with grain yield, which can be used as secondary traits for grain yield selection. Based on linked markers detected and gene action analyzed, an MAS strategy for yield improvement under drought condition could be established, which consists of QTLs contributing to decreased ASI and to increased ear setting and grain yield, respectively.

Detection of Quantitative Trait Loci Associated with Several Internal Organ Traits and Teat Number Trait in a Pig Population

Quantitative trait loci （QTL） were detected for 8 internal organ traits, 3 carcass length traits, and teat number trait in 214 pigs in a resource population that included 180 F2 individuals. A total of 39 microsatellite markers were examined on SSC4, SSC6, SSC7, SSC8, and SSC13. The genetic traits included heart weight （HW）, lung weight （LW）, liver and gallbladder weight （LGW）, spleen weight （SPW）, stomach weight （STW）, small intestine weight （S1W）, large intestine weight （LIW）, kidney weight （KW）, carcass length to the first cervical vertebra （CL1）, carcass length to the first thoracic vertebra （CL2）, rib numbers （RNS）, and teat numbers （TNS）. Results indicated that, 3 highly significant QTL （P≤0.01 at chromosome-wise level） for HW （at 30 cM on SSC6）, RNS （at 115 cM on SSC7）, TNS （at 110 cM on SSC7）, and 6 significant QTL （P≤0.05 at chromosome-wise level） for LW （at 119 cM on SSC13）, LGW （at 94 cM on SSC6）, SPW （at 106 cM on SSC8）, SIW （0 cM on SSC4）, LIW （170 cM on SSC 4）, and TNS （at 95 cM on SSC6） were detected. The phenotypic variances for which these QTL were accounted ranged from 0.04 % to 14.06 %. Most of these QTL had not been previously reported.

Detection of the Quantitative Trait Loci with AFLP and RFLP Markers for Phosphorus Uptake and Use Efficiency in Rice

To identify genetic factors underlying phosphorus (P) uptake and use efficiency under low_P stress in rice (Oryza sativa L.), 84 selected genotypes (recombinant inbred lines) and their parents (which differed in tolerance for low_P stress) “IR20” and IR55178_3B_9_3, were cultured in liquid medium supplemented with adequate and low P levels in a greenhouse. Plants were sampled after 6 weeks in culture for measurements of plant dry weight, P concentration, P uptake and P use efficiency under both P sufficient and stress conditions. A total of 179 molecular markers, including 26 RFLPs and 153 AFLPs, mapped on all 12 chromosomes of rice based on the 84 selected genotypes were used to detect the quantitative trait loci (QTLs) underlying tolerance for low_P stress. Three QTLs were detected on chromosomes 6, 7 and 12, respectively, for relative plant dry weight (RPDW) and relative P uptake (RPUP). One of the QTLs flanked by RG9 and RG241 on chromosome 12 had a major effect which explained about 50% of the variations in the two parameters across the population. The results coincided with the QTLs for low_P stress based on relative tillering ability from the same population from a cross between Nipponbare and Kasalath under soil condition. The identical major QTL for P uptake and plant growth under low_P stress in both liquid medium and soil strongly suggests that the ability of P uptake mainly controls rice tolerance for low_P stress.

Mapping Quantitative Trait Loci Associated with Photoperiod Sensitivity in Maize(Zea mays L.)

Photoperiod sensitivity in maize plays an essential role in utilizing tropic and sub-tropic germplasm to temperate areas. This study aims to identify and map the QTLs responsible for the characteristics measuring photoperiod sensitivity, days from planting to silking （SD）, photoperiod response coefficient of silking （PRC）, and anthesis-silking interval （ASI）. Using the population derived from Zheng 58, photoperiod-insensitive parent, and Ya 8701, photoperiod-sensitive parent, a linkage map was constructed with 93 single sequence repeat （SSR） markers. Phenotyping of 296 F2-3 families of the population in replicated-field test was conducted in both long-day （Beijing, China） and short-day （Sichuan, China） conditions. Ten QTLs were identified to be associated with the SD and ASI on chromosomes 3, 4, 6, 8, and 10 in the longday conditions, and 11 QTLs were detected to be related to the SD and ASI on chromosomes 2, 3, 4, 5, 6, 8, and 10 in the short-day conditions, respectively. A QTL associated with the PRC as a major effect in the long-day conditions located in the same position as the QTL related to the SD and ASI in the map, and was on chromosome 10 linked with marker bnlg1655. Using these QTLs in the marker-assisted selection, the photoperiod sensibility could be reduced by selection of the alleles responsible for the SD, PRC, and ASI in breeding programs.

Quantitative trait loci analysis for root traits in synthetic hexaploid wheat under drought stress conditions

Synthetic hexaploid wheat(SHW),possesses numerous genes for drought that can help breeding for drought-tolerant wheat varieties.We evaluated 10 root traits at seedling stage in 111 F9 recombinant inbred lines derived from a F2 population of a SHW line(SHW-L1)and a common wheat line,under normal(NC)and polyethylene glycol-simulated drought stress conditions(DC).We mapped quantitative trait loci(QTLs)for root traits using an enriched high-density genetic map containing 120370 single nucleotide polymorphisms(SNPs),733 diversity arrays technology markers(DArT)and 119 simple sequence repeats(SSRs).With four replicates per treatment,we identified 19 QTLs for root traits under NC and DC,and 12 of them could be consistently detected with three or four replicates.Two novel QTLs for root fresh weight and root diameter under NC explained 9 and 15.7%of the phenotypic variation respectively,and six novel QTLs for root fresh weight,the ratio of root water loss,total root surface area,number of root tips,and number of root forks under DC explained 8.5–14%of the phenotypic variation.Here seven of eight novel QTLs could be consistently detected with more than three replicates.Results provide essential information for fine-mapping QTLs related to drought tolerance that will facilitate breeding drought-tolerant wheat cultivars.

Mapping of quantitative trait loci using the skew-normal distribution

In standard interval mapping (IM) of quantitative trait loci (QTL), the QTL effect is described by a normal mixture model. When this assumption of normality is violated, the most commonly adopted strategy is to use the previous model after data transformation. However, an appropriate transformation may not exist or may be difficult to find. Also this approach can raise interpretation issues. An interesting alternative is to consider a skew-normal mixture model in standard IM, and the resulting method is here denoted as skew-normal IM. This flexible model that includes the usual symmetric normal distribution as a special case is important, allowing continuous variation from normality to non-normality. In this paper we briefly introduce the main peculiarities of the skew-normal distribution. The maximum likelihood estimates of parameters of the skew-normal distribution are obtained by the expectation-maximization (EM) algorithm. The proposed model is illustrated with real data from an intercross experiment that shows a significant departure from the normality assumption. The performance of the skew-normal IM is assessed via stochastic simulation. The results indicate that the skew-normal IM has higher power for QTL detection and better precision of QTL location as compared to standard IM and nonparametric IM.

Mapping quantitative trait loci associated with starch paste viscosity attributes by using double haploid populations of rice (Oryza sativa L.)

The paste viscosity attributes of starch,measured by rapid visco analyzer(RVA),are important factors for the evaluation of the cooking and eating qualities of rice in breeding programs.To determine the genetic roots of the paste viscosity attributes of rice grains,quantitative trait loci(QTLs)associated with the paste viscosity attributes were mapped,using a double haploid(DH)population derived from Zhongjiazao 17(YK17),a super rice variety,crossed with D50,a tropic japonica variety.Fifty-four QTLs,for seven parameters of the RVA profiles,were identified in three planting seasons.The 54 QTLs were located on all of the 12 chromosomes,with a single QTL explaining 5.99 to 47.11%of phenotypic variation.From the QTLs identified,four were repeatedly detected under three environmental conditions and the other four QTLs were repeated under two environments.Most of the QTLs detected for peak viscosity(PKV),trough viscosity(TV),cool paste viscosity(CPV),breakdown viscosity(BDV),setback viscosity(SBV),and peak time(PeT)were located in the interval of RM 6775-RM 3805 under all three environmental conditions,with the exception of pasting temperature(PaT).For digenic interactions,eight QTLs with six traits were identified for additivexenvironment interactions in all three planting environments.The epistatic interactions were estimated only for PKV,SBV and PaT.The present study will facilitate further understanding of the genetic architecture of eating and cooking quality(ECQ)in the rice quality improvement program.

Quantitative Trait Loci for Resistance to Stripe Disease in Rice(Oryza sativa)

In order to map the quantitative trait loci for rice stripe resistance, a molecular linkage map was constructed based on the F2 population derived from a cross between Zhaiyeqing 8 and Wuyujing 3. Reactions of the two parents, F1 individual and 129 F2：3 lines to, rice stripe were JnvestJgated by both artificial Jnoculation at laboratory and natural infection in the field, and the ratios of disease rating index were scored. The distribution of the ratios of disease rating index in Zhaiyeqing 8/Wuyujing 3 F2：3 population ranged from 0 to 134,08 and from 6.25 to 133.6 under artificial inoculation at laboratory and natural infection in the field, respectively, and showed a marked bias towards resistant parent （Zhaiyeqing 8）, indicating that the resistance to rice stripe was controlled by quantitative trait loci （QTL）. QTL analysis showed that the QTLs detected by the two inoculation methods were completely different. Only one QTL, qSTVT, was detected under artificial inoculation, at which the Zhaiyeqing 8 allele increased the resistance to rice stripe, while two QTLs, qSTV5 and qSTV1, were detected under natural infection, in which resistant alleles came from Zhaiyeqing 8 and Wuyujing 3, respectively. These results showed that resistant parent Zhaiyeqing 8 carried the alleles associated with the resistance to rice stripe virus and the small brown planthopper, and susceptible parent Wuyujing 3 also carried the resistant allele to rice stripe virus. In comparison with the results previously reported, QTLs detected in the study were new resistant genes to rice stripe disease. This will provide a new resistant resource for avoiding genetic vulnerability for single utilization of the resistant gene Stvb-i.

Mapping of Quantitative Trait Loci Affecting Eggshell Quality in an F_2 Population Derived from Strong and Weak Eggshell Lines of the White Leghorn Chicken Breed

Broken and cracked eggshells cause major economic losses to the egg production industry. An F2 population of 262 hens obtained by crossing a strong egg shell line with a weak egg shell line of the White Leghorn breed was used for detecting the quantitative trait loci （QTL） affecting eggshell quality. The 2 lines were developed from the same founder population by two-way selection for egg shell strength with nondestructive deformation. Of the 1 014 microsatellite markers tested, 35 were mapped on 10 autosomal linkage groups. There was no informative marker on chromosome Z. The QTLs associated with 7 traits, i.e., body weight, short length of egg, long length of egg, eggshell strength, eggshell thickness （EST）, eggshell weight （ESW）, and egg weight （EW）, were identified. Highly significant （P〈0.01） QTLs associated with EST and ESW and a significant （P〈 0.05） QTL associated with EW were mapped to a region flanking ABR0545 and ABR0362 on chromosome 9. These QTLs are good candidates to be employed in the development of strategies for reducing the number of broken and cracked eggs in commercial layer houses by employing marker assisted selection.

Quantitative trait loci detection of E dwardsiella tarda resistance in Japanese flounder Paralichthys olivaceus using bulked segregant analysis

In recent years, Edwardsiella tarda has become one of the most deadly pathogens of Japanese fl ounder( Paralichthys olivaceus), causing serious annual losses in commercial production. In contrast to the rapid advances in the aquaculture of P. o livaceus, the study of E. tarda resistance-related markers has lagged behind, hindering the development of a disease-resistant strain. Thus, a marker-trait association analysis was initiated, combining bulked segregant analysis(BSA) and quantitative trait loci(QTL) mapping. Based on 180 microsatellite loci across all chromosomes, 106 individuals from the F1333(♀: F0768 ×♂: F0915)(Nomenclature rule: F+year+family number) were used to detect simple sequence repeats(SSRs) and QTLs associated with E. tarda resistance. After a genomic scan, three markers(Scaffold 404-21589, Scaffold 404-21594 and Scaffold 270-13812) from the same linkage group(LG)-1 exhibited a signifi cant difference between DNA, pooled/bulked from the resistant and susceptible groups( P <0.001). Therefore, 106 individuals were genotyped using all the SSR markers in LG1 by single marker analysis. Two different analytical models were then employed to detect SSR markers with different levels of signifi cance in LG1, where 17 and 18 SSR markers were identifi ed, respectively. Each model found three resistance-related QTLs by composite interval mapping(CIM). These six QTLs, designated q E1–6, explained 16.0%–89.5% of the phenotypic variance. Two of the QTLs, q E-2 and q E-4, were located at the 66.7 c M region, which was considered a major candidate region for E. tarda resistance. This study will provide valuable data for further investigations of E. tarda resistance genes and facilitate the selective breeding of disease-resistant Japanese fl ounder in the future.

Mapping Quantitative Trait Loci Controlling Endosperm Traits with Molecular Marker

Based on the genetic models for triploid endosperm traits and on the methods for mapping diploid quantitative traits loci (QTLs), the genetic constitutions, components of means and genetic variances of QTL controlling endosperm traits under flanking marker genotypes of different generations were presented. From these results, a multiple linear regression method for mapping QTL underlying endosperm traits in cereals was proposed, which used the means of endosperm traits under flanking marker genotypes as a dependent variable, the coefficient of additive effect (d) and dominance effect (h1 and/or h2) of a putative QTL in a given interval as independent variables. This method can work at any position in a genome covered by markers and increase the estimation precision of QTL location and their effects by eliminating the interference of other relative QTLs. This method can also be easily used in other uneven data such as markers and quantitative traits detected or measured in plants and tissues different either in generations or at chromosomal ploidy levels, and in endosperm traits controlled by complicated genetic models considering the effects produced by genotypes of both maternal plants and seeds on them.

Quantitative trait loci associated to agronomic traits and yield components in a Sorghum bicolor L.Moench RIL population cultivated under pre-flowering drought and well-watered conditions

The present study aims to identify QTL influencing agronomic traits and yield components under well-watered and pre-flowering drought stress conditions. One hundred F5 recombinant inbred lines (RIL) and the parental lines of a cross between a drought-tolerant and a susceptible line in a field experiment were carried out at Nong Lam University of Ho Chi Minh City, Vietnam. Drought stress was induced by withholding irrigation water from the plants at four weeks after sowing to flowering. Leaf area of the third leaf, stem diameter, plant height, days to heading, anthesis and maturity, panicle length, number of seeds per plant, hundred kernel weight and grain yield were measured. Plants were genotyped with 117 Diversity Arrays Technology (DArT) and eight expressed sequence tag (EST)-derived simple sequence repeat (SSR) markers. Composite interval mapping was carried out on the traits and significant QTL were claimed at a logarithm of the odds (LOD) score >2.5. A total of 50 QTL were detected on nine chromosomes or 13 linkage groups, respectively. Six promising QTL regions with seven QTL for yield and agronomic traits especially related to pre-flowering drought tolerance were identified on chromosomes SBI-01, SBI-03, SBI-04, SBI-05 and SBI-07.

Mapping Quantitative Trait Loci for Palatability of Milled Rice

Quantitative trait loci (QTLs) controlling palatability in rice were identified using a set of 98 backcross inbred lines (BILs) population derived from a cross between a japonica variety Nipponbare and an indica variety Kasalath. The palatability scores of the population measured by RQ1/Plus Rice Analyzer, showed a continuous and transgressive segregative distribution with a range from 66 to 92. Four putative QTLs for palatability, qPAL-5, qPAL-7, qPAL-8a and qPAL-8b, were detected on chromosome 5, 7 and 8, and they accounted 7.83, 7.03, 11.58 and 7.19% of the total phenotypic variation, respectively. Three alleles qPAL-5, qPAL-7 and qPAL-8b from Kasalath increased the palatability score, whereas only one Nipponbare allele qPAL-8a increased the score . Eight transgressive lines in palatability were selected to make a comparison between phenotypic and genotypic classes. The result explained the possibility of positive QTLs pyramiding through marker-assisted selection of highly palatable rice.

Fine mapping of multiple interacting quantitative trait loci using combined linkage disequilibrium and linkage information

Quantitative trait loci (QTL) and their additive, dominance and epistatic effects play a critical role in complex trait variation. It is often infeasible to detect multiple interacting QTL due to main effects often being confounded by interaction effects. Positioning interacting QTL within a small region is even more difficult. We present a variance component approach nested in an empirical Bayesian method, which simultaneously takes into account additive, dominance and epistatic effects due to multiple interacting QTL. The covariance structure used in the variance component approach is based on combined linkage disequilibrium and linkage (LDL) information. In a simulation study where there are complex epistatic interactions between QTL, it is possible to simultaneously fine map interacting QTL using the proposed approach. The present method combined with LDL information can efficiently detect QTL and their dominance and epistatic effects, making it possible to simultaneously fine map main and epistatic QTL.

Detection of Quantitative Trait Loci Associated with Live Measurement Traits in Pigs

Live measurement growth traits are very important economic traits in pig production and breeding. In this research, quantitative trait loci （QTL） were detected for 11 live estimated growth and carcass traits, including birth weight （BWT）, average daily gain over testing periods （ADG3）, live backfat thickness at last 3-4th lumbar （LBFT3）, live loin eye area （LLEA）, and so on, in 214 pig resource family population, including 180 F2 individual, by 39 microsatellite marker loci on SSC4, SSC6, SSC7, SSC8, and SSC13. The results indicated that 4 chromosome significant level QTL and one suggestive QTL were detected for ADG3 （at position of 50 cM on SSC8）, LBFT3 （at position of 147 cM on SSC4）, LLEA （one highly significant at position of 48 cM on SSC7; another significant at position of 125 cM on SSC8） and BWT （suggestive significant at position of 0 cM, at marker sw489 on SSC4）. The phenotypic variance of these QTL accounted for 0.95% to 16.91%. Most of them were mentioned in previous reports; except the QTL of LLEA at position of sw1953 on SSC8 which maybe a new QTL.

Identification of quantitative trait loci and epistasis for root characteristics and root exudations in maize (Zea mays L.) under deficient phosphorus

The phosphorus uptake （PU） in above-ground parts of plant, root characteristics and root exudations as well as the quantitative trait loci （QTLs） associated with these characteristics were determined for a F2：3 population derived from the cross of two contrasting maize （Zea mays L.） genotypes, 082 and Yel07. A total of 241 F2：3 families were evaluated in replicated trials under deficient phosphorus conditions in 2007 at two sites （Kaixian County and Southwest University, Chongqing, P. R. China）. The results show pleiotropy and close linkage among QTLs. Four common regions in different environments were in bnlg100- bnlg1268b （bins 1.02） for QTL of H＋, bnlg1268a-umc1290a （bins 1.09） for QTL of AP （acid phospbatase activity）, dupssrl5- P 1MT/a （bins 6.06） for QTLs of PU （phosphorus uptake） and RW （root weight）, and P IM3/d-P1M3/g （bins9.04） for QTLs of PU and AP. These QTLs are non-environment or minor QTLs x environment. By epistatic analysis, three main QTLs and eighteen QTLs x QTLs interactions were detected for the seven measured characteristics. These QTLs may affect trait expression by epistatic interaction with the other loci, and make a substantial contribution to phosphorus utilization efficiency, which should be considered when breeding maize varieties with high P efficiency. Two regions were detected in dupssrl 5- P1MT/a （bins 6.06） for QTL of RW and P1M3/d- P 1M3/g （bins 9.04） for QTL of PU and AP. They were detected in two different environments and by two methods of QTL analysis, which were useful for marker-assisted selection.

Dissection of Genetic Effects of Quantitative Trait Loci(QTL) in Transgenic Cotton

When alien DNA inserts into cotton genome in multi-copy manner,several QTL in cotton genome are disrupted,which are called dQTL in this study.Transgenic mutant line is near-isogenic to its recipient which is divergent for the dQTL from remaining QTL.So,a set of data from a

Identification of Quantitative Trait Loci Controlling Seed Physical and Nutrient Traits in Cotton

Cotton(Gossypium spp.) is the leading fiber crop,and an important source of the important edible oil and protein meals in the world.Complex genetics and strong environmental effects hinder

Identification of quantitative trait loci for growth-related traits in the blood clam(Tegillarca granosa)

The blood clam(Tegillarca granosa)is an economically important marine shellfish in the southeast coastal area of China.In T.granosa breeding practice,growth-related traits are most important phenotypic selection index.In order to increase the breeding efficiency,we used a previously published genetic linkage map of T.granosa to map quantitative trait loci(QTL)for growth-related traits in a single full sibling F1 family.Eight growth-related traits including shell length(SL),shell height(SH),shell width(SW),shell weight(Swi),shell umbo width(SUW),as well as outer ligament length(OLL),meat weight(MW)and total weight(TW)of T.granosa were measured for QTL mapping.The estimated phenotypic correlations between any 2 of the 8 growth-related traits were significant(P<0.001).Twenty-nine QTL were identified for growth-related traits,including 13 QTL with the phenotypic variance of 4.20-15.63%in the female map and 16 QTL of 3.99%-14.58%in the male map.Thirteen amplification fragment length polymorphism(AFLP)markers were validated to closely link to seven QTL except the shell height.This is the first report of QTL mapping on growth-related traits in T.granosa.These QTL and their tightly linked markers present the initial step towards the goal of marker-assisted selection in T.granosa.