Genetic analysis of sugar-related traits in rice grain

Sugar is the primary product of photosynthesis in plant and plays a critical role in regulating plant growth and development. In this study, quantitative trait loci (QTLs) for total soluble sugar, sucrose, and fructose contents in rice grain were identified using a double haploid population derived from a cross between japonica CJ06 and indica TN1. A total of 17 QTLs, including four QTLs for total soluble sugar content, seven QTLs for sucrose content, and six QTLs for fructose content, were detected on chromosome 1, 3, 4, 5, 6, and 8, with the LOD ranges from 2.61 to 3.85. Furthermore, among the determined varieties, we found that the total soluble sugar content in japonica showed higher than that in indica. Comparative genetic analysis showed that starch synthesis related gene is presumably involved in sugar-related metabolic activity in rice grain.     

Genetic and molecular dissection of quantitative traits in rice

Recent progress in the generation of a molecular genetic map and markers for rice has made possible a new phase of mapping individual genes associated with complex traits. This type of analysis is often referred to as quantitative trait locus (QTL) analysis. Increasing numbers of QTL analyses are providing enormous amounts of information about QTLs, such as the numbers of loci involved, their chromosomal locations and gene effects. Clarification of genetic bases of complex traits has a big impact not only on fundamental research on rice plant development, but it also has practical benefits for rice breeding. In this review, we summarize recent progress of QTL analysis of several complex traits in rice. A strategy for positional cloning of genes at QTLs is also discussed.     

Genetic dissection of sorghum grain quality traits using diverse and segregating populations

Key message

Coordinated association and linkage mapping identified 25 grain quality QTLs in multiple environments, and fine mapping of the Wx locus supports the use of high-density genetic markers in linkage mapping.

Abstract

There is a wide range of end-use products made from cereal grains, and these products often demand different grain characteristics. Fortunately, cereal crop species including sorghum [Sorghum bicolor (L.) Moench] contain high phenotypic variation for traits influencing grain quality. Identifying genetic variants underlying this phenotypic variation allows plant breeders to develop genotypes with grain attributes optimized for their intended usage. Multiple sorghum mapping populations were rigorously phenotyped across two environments (SC Coastal Plain and Central TX) in 2 years for five major grain quality traits: amylose, starch, crude protein, crude fat, and gross energy. Coordinated association and linkage mapping revealed several robust QTLs that make prime targets to improve grain quality for food, feed, and fuel products. Although the amylose QTL interval spanned many megabases, the marker with greatest significance was located just 12 kb from waxy (Wx), the primary gene regulating amylose production in cereal grains. This suggests higher resolution mapping in recombinant inbred line (RIL) populations can be obtained when genotyped at a high marker density. The major QTL for crude fat content, identified in both a RIL population and grain sorghum diversity panel, encompassed the DGAT1 locus, a critical gene involved in maize lipid biosynthesis. Another QTL on chromosome 1 was consistently mapped in both RIL populations for multiple grain quality traits including starch, crude protein, and gross energy. Collectively, these genetic regions offer excellent opportunities to manipulate grain composition and set up future studies for gene validation.

     

Molecular dissection of QTL governing grain size traits employing association and linkage mapping in Basmati rice

Grain size is one of the key traits that determines the quality of Basmati rice from the consumers’ as well as the traders’ point of view. Though many genes governing grain size have been identified in indica and japonica, little work has been done in Basmati rice. The present study aims at dissection of a QTL region governing grain size traits in Basmati employing association and linkage mapping approaches. Association mapping revealed that three markers, i.e., RM 6024 (grain breadth), RM1237 and RM18582 (grain length-breadth ratio), which cover 889 kb in the targeted QTL region have been significantly associated with grain size traits. Using linkage mapping, the targeted QTL region has been further delimited to a physical distance of 268 kb that comprises 24 annotated genes. The gene expression analysis of parents, revealed 19 genes differentially expressing within the QTL. Of them, 15 genes showed high expression in Basmati370, while four were expressed in Jaya, and whereas five genes did not show any differential expression between parents. Among differentially expressed genes, a highly expressed gene in Basmati370, Os05g0374200 (Cytokinin dehydrogenase 1 precursor) seems to be involved in accumulation of cytokinins, thus affecting the grain size. Therefore, our findings demonstrated that by complimenting association and linkage mapping, it is likely to dissect a QTL governing grain size traits in Basmati rice and also the QTL could be a potential target for marker-assisted breeding and map-based cloning studies.     

Genetic dissection of the resistance to Rice stripe virus present in the indica rice cultivar 'IR24'

Rice stripe disease, caused by Rice stripe virus (RSV) and transmitted by the small brown planthopper (Laodelphax striatellus Fallen), is one of the most serious viral diseases of rice in temperate East Asian production regions. Prior quantitative trait loci (QTL) mapping has established that Oryza sativa L. subsp. indica 'IR24' carries positive alleles at the three loci qSTV3, qSTV7, and qSTV11-i. Here, we report an advanced backcross analysis based on three selected chromosome segment substitution lines (CSSLs), each predicted to carry one of these three QTL. Three sets of BC(4)F(2:3) populations were bred from a cross between the critical CSSL and its recurrent parent Oryza sativa L. subsp. japonica 'Asominori'. Both qSTV3 and qSTV11-i were detected in their respective population, but qSTV7 was not. An allelic analysis based on a known carrier of the major RSV resistance gene Stvb-i, which is located on chromosome 11, showed that qSTV11-i was not allelic with Stvb-i. A large mapping population was used to delimit the location of qSTV11-i to a 73.6-kb region. The de novo markers developed for this purpose will be useful as marker-assisted selection tools in efforts to introduce qSTV11-i into breeding programmes aiming to improve the level of RSV resistance.     

Genetic analysis of rice grain quality

 The inheritance of grain quality is more complicated than that of other agronomic traits in cereals due to epistasis, maternal and cytoplasmic effects, and the triploid nature of endosperm. In the present study, an established rice DH population derived from anther culture of an indica/japonica hybrid was used for genetic analysis of rice grain quality. A total of five parameters, amylose content (AC), alkali-spreading score (ASS), gel consistency (GC), percentage of grain with a white core (PGWC) and the square of the white core (SWC), were estimated for the DH lines and the parent varieties. For each parent, the value of each parameter was relatively stable in three locations, Beijing, Hangzhou and Chengdu, while the differences between the parents were significant for all five parameters. AC showed a bimodal distribution, and the distribution of ASS was skewed toward the value of JX17, while the other three parameters displayed continuous distributions among the DH lines with partially transgressive segregations. For AC, a minor and a major gene were found on chromosomes 5 and 6 respectively. The major gene, which should be an allele of wx, explained 91.9% of the total variation. For GC, two QTLs were identified on chromosomes 2 and 7 respectively. For ASS, a minor and a major gene were both located on chromosome 6. The major gene should be the same locus as the alkali degeneration gene (alk). Genetic linkage between alk and wx was found in QTL mapping. For PGWC, two QTLs were located on chromosomes 8 and 12. Only a minor QTL was found for SWC on chromosome 3. The results and the molecular markers presented here may be useful in rice breeding for grain quality improvement. Received: 24 April 1998 / Accepted: 13 August 1998     

Molecular dissection of seedling-vigor and associated physiological traits in rice

seedling-vigor is important for crop establishment. There have been reported quantitative trait locus (QTL) analyses on seedling-vigor related morphological traits. However, physiological understanding of these detected QTLs is rather limited. In this study, we employed a recombinant inbred population to detect QTLs for seedling-vigor traits and physiological traits related to seedling-vigor. Germination rate and seedling growth were measured to quantify seedling-vigor. Total amylase activity, !-amylase activity, reducing sugar content, root activity and seed weight were determined. Correlations were observed between the seedling-vigor and physiological traits. QTL analysis reveals that the intervals of RG393-C1087-RZ403 on chromosome 3, C246-RM26-C1447 and R830-R3166-RG360-C734b on chromosome 5, and the interval of Waxy on chromosome 6 are the four main chromosomal regions controlling seedling-vigor. Several QTLs for amylase activities, reducing sugar content and root activity were localized in the similar regions as the QTLs for seedling-vigor. The results suggest that these traits were under the control of pleiotropic and/or closely linked QTLs. The implications of the results in the understanding of the physiological basis of seedling-vigor were discussed.     

Genetic dissection and pyramiding of quantitative traits for panicle architecture by using chromosomal segment substitution lines in rice

To understand the genetic basis of yield-related traits of rice, we developed 39 chromosome segment substitution lines (CSSLs) from a cross between an average-yielding japonica cultivar, Sasanishiki, as the recurrent parent and a high-yielding indica cultivar, Habataki, as the donor. Five morphological components of panicle architecture in the CSSLs were evaluated in 2 years, and 38 quantitative trait loci (QTLs) distributed on 11 chromosomes were detected. The additive effect of each QTL was relatively small, suggesting that none of the QTLs could explain much of the phenotypic difference in sink size between Sasanishiki and Habataki. We developed nearly isogenic lines for two major QTLs, qSBN1 (for secondary branch number on chromosome 1) and qPBN6 (for primary branch number on chromosome 6), and a line containing both. Phenotypic analysis of these lines revealed that qSBN1 and qPBN6 contributed independently to sink size and that the combined line produced more spikelets. This suggests that the cumulative effects of QTLs distributed throughout the genome form the major genetic basis of panicle architecture in rice. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. T. Ando and T. Yamamoto contributed equally to this work.     

Genetic basis of 17 traits and viscosity parameters characterizing the eating and cooking quality of rice grain

A recombinant inbred line population derived from a cross between Zhenshan 97 and Delong 208 was used to analyze the genetic basis of the cooking and eating quality of rice as reflected by 17 traits (or parameters). These traits include amylose content (AC), gel consistency (GC), alkali spreading value (ASV), cooked rice elongation (CRE), and 13 parameters from the viscosity profile. All the traits, except peak paste viscosity (PKV), time needed from gelatinization to peak (BAtime), and CRE, can be divided into two classes according to their interrelationship. The first class consists of AC, GC, and most of the paste viscosity parameters that form a major determinant of eating quality. The second class includes ASV, pasting temperature (Atemp) and pasting time (Atime), which characterize cooking process. We identified 26 QTL (quantitative trait locus or loci) in 2 years; nine QTL clusters emerged. The two major clusters, which correspond to the Wx and Alk loci, control the traits in the first and second classes, respectively. Some QTL are co-located for the traits belonging to the same class and also for the traits to a different class. The Wx locus also affects on ASV while the Alk locus also makes minor contributions to GC and some paste viscosity parameters. The QTL clusters on other chromosomes are similar to the Wx locus or Alk locus, although the variations they explained are relatively minor. QTL for CRE and PKV are dispersed and independent of the Wx locus. Low paste viscosity corresponds to low AC and soft gel, which represents good eating quality for most Chinese consumers; high ASV and low Atemp, together with reduced time to gelatinization and PKV, indicate preferred cooking quality. The genetic basis of Atemp, Atime, BAtime, peak temperature, peak time, paste viscosity at 95 degrees C, and final paste viscosity is newly examined to reveal a complete and dynamic viscosity profile.     

Genetic dissection of complex traits using hierarchical biological knowledge

Despite the growing constellation of genetic loci linked to common traits, these loci have yet to account for most heritable variation, and most act through poorly understood mechanisms. Recent machine learning (ML) systems have used hierarchical biological knowledge to associate genetic mutations with phenotypic outcomes, yielding substantial predictive power and mechanistic insight. Here, we use an ontology-guided ML system to map single nucleotide variants (SNVs) focusing on 6 classic phenotypic traits in natural yeast populations. The 29 identified loci are largely novel and account for ~17% of the phenotypic variance, versus <3% for standard genetic analysis. Representative results show that sensitivity to hydroxyurea is linked to SNVs in two alternative purine biosynthesis pathways, and that sensitivity to copper arises through failure to detoxify reactive oxygen species in fatty acid metabolism. This work demonstrates a knowledge-based approach to amplifying and interpreting signals in population genetic studies.     

Genetic diversity and phylogeny of Japanese sake-brewing rice as revealed by AFLP and nuclear and chloroplast SSR markers

Japanese rice (Oryza sativa L.) cultivars that are strictly used for the brewing of sake (Japanese rice wine) represent a unique and traditional group. These cultivars are characterized by common traits such as large grain size with low protein content and a large, central white-core structure. To understand the genetic diversity and phylogenetic characteristics of sake-brewing rice, we performed amplified fragment length polymorphism and simple sequence repeat analyses, using 95 cultivars of local and modern sake-brewing rice together with 76 cultivars of local and modern cooking rice. Our analysis of both nuclear and chloroplast genome polymorphisms showed that the genetic diversity in sake-brewing rice cultivars was much smaller than the diversity found in cooking rice cultivars. Interestingly, the genetic diversity within the modern sake-brewing cultivars was about twofold higher than the diversity within the local sake-brewing cultivars, which was in contrast to the cooking cultivars. This is most likely due to introgression of the modern cooking cultivars into the modern sake-brewing cultivars through breeding practices. Cluster analysis and chloroplast haplotype analysis suggested that the local sake-brewing cultivars originated monophyletically in the western regions of Japan. Analysis of variance tests showed that several markers were significantly associated with sake-brewing traits, particularly with the large white-core structure.     

Genetic dissection of floral traits in anemone-type chrysanthemum by QTL mapping

Brassica oleracea comprises several important subspecies, including cabbage, broccoli, cauliflower, Chinese kale, and kohlrabi. The petal color of Chinese kale is mostly white and sometimes yellow. To explore the genetic basis of petal color variation in Chinese kale, F₂ and BC₁ (backcross) populations were constructed from the cross of two inbred lines, 2114 (yellow petal) and 2116 (white petal). Genetic analysis of the F₂ and BC₁ populations demonstrated that yellow petal color was controlled by a single recessive nuclear gene, termed cpc-2. Insertion-deletion (InDel) markers, designed based on the parental resequencing data, were used to map cpc-2. The fine mapping results indicated that the cpc-2 gene was located in a 569-kb interval on chromosome C03 flanked by InDel markers ZB636 and ZB692, with genetic distances of 0.3 cM and 0.6 cM, respectively. By analyzing the nucleotide variations and annotations of the genes in this interval, a CCD4 family gene was predicted to be a candidate for cpc-2 and renamed BoCCD4.2. In addition, insertion of the CACTA-like transposable element (TE3) interrupted the function of the BoCCD4 gene, which may have resulted in the loss of function of BoCCD4 and the petal color transition from white to yellow. The TE3 insertion in the BoCCD4 gene was also present in 63 cabbage inbred lines among 159 accessions, which revealed that the TE3-type null allele of BoCCD4 formed before the divergence of the two subspecies cabbage and Chinese kale and that Chinese kale evolved much earlier than cabbage. This study lays the foundation for cloning BoCCD4.2 and revealing the molecular mechanism underlying petal color formation in Chinese kale.

     

Genetic dissection of heading time and its components in rice

Heading time (HT, days from sowing to heading) is an important agronomic trait in rice. Physiologically, HT can be divided into two stages: vegetative growth time (VGT) and reproductive growth time (RGT). A number of studies for mapping QTLs conferring HT in rice have been reported, but none of them has tried to map HT-related QTLs based on their component traits (VGT and RGT). The present study aims to map HT-related QTLs in rice according not only to the performance of HT, but also the performances of VGT and RGT. A method based on an empirical equation of leaf age growth was developed to partition HT into VGT and RGT. An indica/japonica DH population and a corresponding RFLP map were constructed for the study. The methods of composite interval mapping and multiple-trait composite interval mapping were used to map QTLs. A total of 19 QTLs were mapped on all 12 rice chromosomes with the exception of chromosomes 1 and 4. Results showed that: (1) more QTLs could be detected by partitioning HT into VGT and RGT; (2) the genetic variation of HT was largely attributed to VGT; and (3) the two component stages were relatively independent in terms of QTL effects, suggesting that the ratio between VGT and RGT could be genetically adjusted without apparently altering HT. Received: 1 June 2000 / Accepted: 16 October 2000     

Validation of gene based marker-QTL association for grain dimension traits in rice

Validation of marker-QTL association for genes grain size 3 (GS3), grain weight 2 (GW2), seed width 5 (qSW5) and a QTL qgrl7.1 for grain length was undertaken in a set of 242 diverse rice germplasm. Further, the study was extended to an F₂ mapping population derived from cross of Sonasal, a short grain aromatic rice landrace with Pusa Basmati 1121, a variety with extra long slender grains. Seven gene specific markers, namely, SF28, SR17, RGS1and RGS2 based on GS3, W004 for GW2, MS40671 for qSW5 and RM505 for qgrl7.1, were used for validation. Single marker analysis revealed significant association of these markers to grain size and shape. The marker SF28 explained highest phenotypic variance (37 %) while the marker W004 explained lowest variance (2.6 %) for grain length in the germplasm set at the significance level P?<?0.05. Three markers namely, SF28, MS40671 and RM505 were polymorphic between the parents Sonasal and Pusa Basmati 1121. In the F₂ population, the marker SF28 linked to gene GS3 explained highest phenotypic variance (32.5 %), while RM505 linked to qgrl7.1 explained 5.4 % of phenotypic variance for grain length. The marker SF28 was found to be most robust in the validation studies both in germplasm and F₂ population. The validated gene specific markers can be utilised in marker assisted selection for improving grain size and shape as these traits have significant contribution towards grain quality and grain yield. This is the first study on validation of gene based markers for grain dimension traits in Indian rice germplasm.     

Dissecting the Genetic Basis of Extremely Large Grain Shape in Rice Cultivar ＇JZ1560＇

Rice grain shape,grain length(GL),width(GW),thickness(GT)and length-to-width ratio(LWR),are usually controlled by multiple quantitative trait locus(QTL).To elucidate the genetic basis of extremely large grain shape,QTL analysis was performed using an F2 population derived from a cross between a japonica cultivar ’JZ1560’(extremely large grain)and a contrasting indica cultivar ’FAZ1’(small grain).A total number of 24 QTLs were detected on seven different chromosomes.QTLs for GL,GW,GT and LWR explained 11.6%,95.62%,91.5%and 89.9%of total phenotypic variation,respectively.Many QTLs pleiotropically controlled different grain traits,contributing complex traits correlation.GW2 and qSW5/GW5,which have been cloned previously to control GW,showed similar chromosomal locations with qGW2-I/qGT2-I/qLWR2-2 and qGW5-2/qLWR5-l and should be the right candidate genes.Plants pyramiding GW2 and qSW5/GW5 showed a significant increase in GW compared with those carrying one of the two major QTLs.Furthermore,no significant QTL interaction was observed between GW2 and qSW5/GW5.These results suggested that GW2 and qSW5/GW5 might work in independent pathways to regulate grain traits.’JZ1560’ alleles underlying all QTLs contributed an increase in GW and GT and the accumulation of additive effects generates the extremely large grain shape in ’JZ1560’.     

稻米品质性状对开放式空气二氧化碳浓度增高的响应

利用开放式空气CO2浓度增高(FACE)系统平台。研究大田栽培条件下粳稻武香粳14号稻米品质性状对CO2浓度增高200μmol·mol^-1的响应。结果表明．FACE处理稻谷的出糙率平均比CK高1．4个百分点,整精米率平均比CK低12．3个百分点,较低的供N水平有利于提高FACE条件下的出糙率．较高的供N水平有利于提高FACE条件下的整精米率;FACE处理的稻米垩白略有增加。垩白粒率平均比CK高11．9个百分点,垩白度平均比CK平均高2．8个百分点,较高的供N和供P水平有利于降低FACE条件下垩白大小、垩白粒率和垩白度;FACE处理稻米糊化温度平均比CK平均高0．52℃,胶稠度有提高的趋势,但对稻米直链淀粉含量影响较小,较高的供N和供P水平有利于降低FACE条件下稻米的直链淀粉含量,较低的供N和较高的供P水平有利于降低FACE条件下稻米胶稠度,较低的供N水平有利于降低FACE条件下稻米糊化温度;FACE处理使稻米蛋白质含量比CK平均低0．6个百分点,较低的供N和供P水平有利于降低FACE条件下稻米蛋白质含量。     

Genetic dissection of photosynthetic efficiency traits for enhancing seed yield in chickpea

Understanding the genetic basis of photosynthetic efficiency (PE) contributing to enhanced seed yield per plant (SYP) is vital for genomics‐assisted crop improvement of chickpea. The current study employed an integrated genomic strategy involving photosynthesis pathway gene‐based association mapping, genome‐wide association study, quantitative trait loci (QTL) mapping, and expression profiling. This identified 16 potential single nucleotide polymorphism loci linked to major QTLs underlying 16 candidate genes significantly associated with PE and SYP traits in chickpea. The allelic variants were tightly linked to positively interacting QTLs regulating both enhanced PE and SYP traits as exemplified by a chlorophyll A‐B binding protein‐coding gene. The leaf tissue‐specific pronounced up‐regulated expression of 16 associated genes in germplasm accessions and homozygous individuals of mapping population was evident. Such combinatorial genomic strategy coupled with gene haplotype‐specific association and in silico protein–protein interaction study delineated natural alleles and superior haplotypes from a chlorophyll A‐B binding (CAB) protein‐coding gene and its interacting gene, Timing of CAB Expression 1 (TOC1), which appear to be most promising candidates in modulating chickpea PE and SYP traits. These functionally pertinent molecular signatures identified have efficacy to drive marker‐assisted selection for developing PE‐enriched cultivars with high seed yield in chickpea.