MAGIC群体的遗传特征及其在作物耐逆研究上的应用

多亲本高世代互交（multi-parent advanced generation intercross,MAGIC）群体是近年来发展起来的新一代遗传作图及育种群体。MAGIC群体最初是以研究动物及人类复杂性状遗传基础为目标而构建的基于多亲本的重组近交群体,随后将其构建方法衍生到植物中应用。MAGIC群体应用于作物遗传育种,可以建立包含无限多株系的群体,主要的优势是拥有大量可利用的多样性遗传基因池。亲本可选用育种中性状优异的材料,通过多次重组创造大量的遗传变异。群体中选出的优良株系可用作育种中间材料或直接组配新品种,也可灵活应用于数量性状位点（quantitative trait locus,QTL）的精确遗传定位分析,真正做到育种群体和定位群体的有机整合。作物耐逆性大多是由QTL控制,因此,主要围绕MAGIC群体的定义、构建流程、遗传特征及其在作物耐逆性研究上的应用和发展前景作综合性阐述。     

大豆苗期耐低磷性及其QTL定位

利用来自波高和南农94-156（耐低磷种质）的重组自交系群体NJ(SP)BN（151个家系）通过盆栽试验研究与耐低磷有关的性状,并进行耐低磷性状的QTL定位。初步结果表明,不施磷处理的总干重主要由单株P吸收量决定,而与磷利用效率无关;而单株P吸收量与根干重、根效率均极显著正相关,单株P吸收量变异的76.2%由根效率决定。不施磷处理的根冠比（R/S）显著增加主要是茎干重无显著变化而根干重显著增加所致。在D1b+W、F、G、N和O等5个连锁群上共检测到7个QTL与耐低磷有关。分别可解释所对应性状表型变异的4.8%~17.0%,其中5个QTL的增效基因来自亲本波高,2个QTL的增效基因来自亲本南农94-156。     

黄淮海地区大豆品种苗期根系性状的遗传变异及与耐逆境胁迫的关系

从83份黄淮海地区代表性大豆地方品种和育成品种(系)中按根系类型选取32份,用以研究苗期根系性状的遗传特点、与地上部性状的相关以及与逆境胁迫的关系。大豆苗期一级侧根数、主根长、根干质量、总根长和根体积等性状,在品种间、各苗龄间均存在显著遗传变异;根系性状与整株干质量呈高度相关;根干质量、根总长和根体积的相对值与耐旱平均隶属函数值,一级侧根数、主根长、总根长、根体积、根干质量的相对值与耐铝毒平均隶属函数值呈极显著相关,且根系性状的相对值在品种间存在显著变异,可用做耐逆性选择的根系指标。     

Quantitative trait loci mapping for flooding tolerance at an early growth stage of soybean recombinant inbred line population

Flooding stress causes a significant yield reduction in soybean. The early growth of soybean in Korea coincides with the rainy season, potentially exposing to flooding stress. Greenhouse experiments were conducted to map the quantitative trait loci (QTL) for flooding tolerance in soybean and to identify and investigate candidate genes near the QTL hot spots. Flood stress was imposed at V1–V2 stage on a recombinant inbred line population (‘Paldalkong’ × ‘NTS1116’), and leaf chlorophyll content (CC) and shoot dry weight (DW) were measured under control and flooded conditions. The genetic map was constructed using 180K Axiom^® SoyaSNP markers. The QTL were analysed under control and flooded conditions as well as for index (ratio of CC or DW under flooded to control, CCI and DWI) and flooding tolerance index (FTI, mean of CCI and DWI). A total of 20 QTL with LOD scores 3.59–19.73 causing 5.8%–33.3% phenotypic variation were identified on nine chromosomes. Chromosomes 10, 12 and 13 harboured relatively more stable QTL. Results of this study could be useful to further understand the genetic basis of soybean's flooding tolerance and applied in breeding programmes.     

QTL mapping of phosphorus deficiency tolerance in soybean (Glycine max L. Merr.)

Phosphorus (P) deficiency is a major abiotic stress that limits plant growth and crop productivity throughout the world. In the present study, 184 recombinant inbred line (RIL) families developed from soybean varieties Kefeng No. 1 and Nanong 1138-2 were used to identify quantitative trait loci (QTL) associated with P deficiency tolerance. Seven traits of plant height (HT), weight of fresh shoot (FSW), weight of fresh root (FRW), weight of dry root (DRW), length of main root (RL), phosphorus content in leaf (LP), phosphorus content in root (RP), were used as parameters to assess the phosphorus deficiency tolerance. The QTL mapping for the seven traits was performed using the program WinQTLCart. Seven QTLs were detected and mapped on two linkage groups for three traits of weight of fresh shoot, phosphorus contents in leaf and in root. The QTLs that had LOD scores more than three were detected for all of the three traits above. Most of the QTLs explained more than 10% of the total variation. The two QTLs for phosphorus content in leaf explained more than 20% of the total variation, respectively. Five QTLs were mapped on linkage group F2, and two on linkage F1. It was suggested that the genes related to phosphorus deficiency tolerance located on linkage group F in soybean.Contributed equally to this work.     

Association analysis for detecting significant single nucleotide polymorphisms for phosphorus-deficiency tolerance at the seedling stage in soybean [Glycine max (L) Merr.]

Tolerance to low-phosphorus soil is a desirable trait in soybean cultivars. Previous quantitative trait locus (QTL) studies for phosphorus-deficiency tolerance were mainly derived from bi-parental segregating populations and few reports from natural population. The objective of this study was to detect QTLs that regulate phosphorus-deficiency tolerance in soybean using association mapping approach. Phosphorus-deficiency tolerance was evaluated according to five traits (plant shoot height, shoot dry weight, phosphorus concentration, phosphorus acquisition efficiency and use efficiency) comprising a conditional phenotype at the seedling stage. Association mapping of the conditional phenotype detected 19 SNPs including 13 SNPs that were significantly associated with the five traits across two years. A novel cluster of SNPs, including three SNPs that consistently showed significant effects over two years, that associated with more than one trait was detected on chromosome 3. All favorable alleles, which were determined based on the mean of conditional phenotypic values of each trait over the two years, could be pyramided into one cultivar through parental cross combination. The best three cross combinations were predicted with the aim of simultaneously improving phosphorus acquisition efficiency and use efficiency. These results will provide a thorough understanding of the genetic basis of phosphorus deficiency tolerance in soybean.     

Genetic variation for maize root architecture in response to drought stress at the seedling stage

Although the root system is indispensable for absorption of nutrients and water, it is poorly studied in maize owing to the difficulties of direct measurement of roots. Here, 103 maize lines were used to compare root architectures under well-watered and water-stressed conditions. Significant genetic variation, with medium to high heritability and significant correlations, was observed for root traits. Total root length (TRL) and total root surface area (TSA) had high phenotypical diversity, and TRL was positively correlated with TSA, root volume, and root forks. The first two principal components explained 94.01% and 91.15% of total root variation in well-watered and water-stressed conditions, respectively. Thus, TRL and TSA, major contributors to root variation, can be used as favorable selection criteria at the seedling stage. We found that stiff stalk and non-stiff stalk groups (temperate backgrounds) showed relatively higher mean values for root morphological diversity than the TST group (tropical/subtropical background). Of the tested lines, 7, 42, 45, and 9 were classified as drought sensitive, moderately sensitive, moderately drought tolerant, and highly drought tolerant, respectively. Seven of the 9 extremely drought tolerant lines were from the TST group, suggesting that TST germplasms harbor valuable genetic resources for drought tolerance that could be used in breeding to improve abiotic stress tolerance in maize.     

A major QTL associated with cold tolerance at seedling stage in rice (Oryza sativa L.)

Chilling injury is one of the most important limiting factors affecting rice production in temperate and high-elevation areas. In this study, 146 microsatellite markers were employed to identify quantitative trait loci (QTL) conferring cold tolerance at seedling stage (CTS) .The mapping population consisted of 193 doubled haploid (DH) lines, which derived from a cross between a cold-tolerant japonica variety (AAV002863) and a cold-sensitive indica cultivar (Zhenshan97B). Tolerance to cold was assessed by the survival percentage of seedlings after cold treatment. In a climate chamber, after treatment at 6°C/10°C for 7 d, the measurement was taken on the sixth day of the recovery stage at room temperature. The phenotypic distribution of the DH population approximately fitted normality with skewness and kurtosis less than 0.3, and the difference among the three repetitions was not significant. Five main effect QTLs were identified with LOD > 4.0 on chromosomes 1, 2, 8 using a composite interval mapping approach. The accumulated contribution of the five QTLs was 62.28%, and a major QTL (LOD = 15.09) was identified on chromosome 2 flanked by RM561 and RM341, which explained 27.42% of the total phenotypic variation. Four significant epistatic interactions were also detected with a total contribution of 20.14%. Liang Chen and Qiaojun Lou had made the equal contribution for the research.     

QTL analysis of flooding tolerance in soybean at an early vegetative growth stage

Soybean cultivars are sensitive to flooding stress and their seed yields are substantially reduced in response to the stress. This study was conducted to investigate the genetic basis of flooding tolerance at an early vegetative growth stage. Sixty recombinant inbred lines derived from a cross between a relatively tolerant cv. ‘Misuzudaizu’ and a sensitive cv. ‘Moshidou Gong 503’ were grown in pots in a vinyl plastic greenhouse in 2002 and 2003. At the two‐leaf stage, half of the pots were waterlogged by water placed in plastic containers and adjusted to 5 cm above the soil surface. After 3 weeks of treatment, the pots were returned to the greenhouse and grown until maturity. Flooding tolerance was evaluated by dividing the seed weight of the treated plants by that of the control plants. Quantitative trait loci (QTL) analysis using 360 genetic markers revealed three QTLs for flooding tolerance, ft1 to ft3 in 2002. The ft1 (molecular linkage group C2) was reproducible and an additional four QTLs, ft4 to ft7, were found in 2003. The ft1 had a high LOD score in both years (15.41 and 7.57) and accounted for 49.2% and 30.5% of the total variance, respectively. A large QTL for days to flowering was consistently observed across treatments and years at a similar position to ft1. Comparing the relative location with markers, the maturity gene probably corresponds to E1. Late maturity may have conferred a longer growth period for recovery from flooding stress.     

QTL pyramiding for improving of cold tolerance at fertilization stage in rice

Vigorous cold tolerance at the fertilization stage (CTF) is a very important characteristic for stable rice production in cold temperature conditions. Because CTF is a quantitatively inherited trait, pyramiding quantitative trait loci (QTLs) using marker-assisted selection (MAS) is effective for improving CTF levels in rice breeding programs. We previously identified three QTLs controlling CTF, qCTF7, qCTF8 and qCTF12, using backcrossed inbred lines derived from a cross between rice cultivar Eikei88223 (vigorous CTF) and Suisei (very weak CTF). However, pyramiding of these QTLs for the application of MAS in practical rice breeding programs have not yet been elucidated. In this study, we examined the effect of pyramiding QTLs for improvement of CTF level using eight possible genotype classes from the 152 F₃ population derived from a cross between Eikei88223 and Suisei. Increasing of CTF levels in combinations between qCTF7 and qCTF12 and between qCTF8 and qCTF12 were detected. Furthermore, we compared the haplotype pattern around the QTLs for CTF among the rice cultivars from Hokkaido. These results are useful for improvement of new cultivars with high CTF levels using MAS and identification of genetic resources with the novel QTL(s) for CTF.     

Detection of quantitative trait loci for phosphorus deficiency tolerance at soybean seedling stage

Phosphorus deficiency is a primary constraint to soybean productivity in acid and calcareous soils. Our aim was to map quantitative trait loci (QTL) controlling phosphorus deficiency tolerance using 152 recombinant inbred lines derived from a cross between the P stress tolerant variety Nannong94-156 and the P stress sensitive variety Bogao. Five traits were used as parameters to evaluate phosphorus deficiency tolerance at seedling stage under different phosphorus levels in experiments 2005 and 2006. As a result, thirty-four additive QTLs were detected on nine linkage groups, with corresponding contribution ratios of 6.6–19.3%. There were three clusters of QTL found in genomic regions S506-Satt534 (on linkage group B2-1), Sat_183-Satt274 (on linkage group D1b + W), and Sat_185-Satt012 (on linkage group G). The locus flanked by Sat_183-Satt274 on linkage group D1b + W was coincident with four previously discovered QTLs with phosphorus efficiency. Another interesting locus flanked by Sat_185-Satt012 on linkage group G was detected across years. The identified QTL will be useful to improve the stress resistance of soybean against a complex nutritional disorder caused by phosphorus deficiency. In addition, more QTLs were detected under low phosphorus condition and some QTLs were detected that specifically expressed under different phosphorus levels. These particular QTLs could help provide greater understanding of the genetic basis of phosphorus efficiency in soybean.     

Identification of QTLs controlling rice drought tolerance at seedling stage in hydroponic culture

‘Drought avoidance’ and ‘drought tolerance’ are two mechanisms by which plants adapt under water stress. These mechanisms are difficult to evaluate separately in field experiments. Using hydroponic culture, we studied the genetic control of drought tolerance in rice (Oryza sativa L.) without the effect of drought avoidance. A backcross inbred population of ‘Akihikari’ (lowland cultivar) × ‘IRAT109’ (upland cultivar) with 106 lines was cultured with (stressed condition) and without (non-stressed condition) polyethylene glycol (PEG) at seedling stage. The relative growth rate (RGR), specific water use (SWU), and water use efficiency (WUE) showed significant genotype × environment interactions with or without PEG, indicating that each line responded differently to water stress. A quantitative trait locus (QTL) analysis revealed that these interactions were QTL specific. A total of three QTLs on chromosomes 2, 4, and 7 were detected for RGR. The QTL on chromosome 7 had a constant effect across environments, while the QTL on chromosome 4 had an effect only under non-stressed condition and that on chromosome 2 only under stressed condition. The stress-specific QTL on chromosome 2 was not co-located with any QTLs for root system depth previously reported from the same mapping population. However, this QTL was co-located with a stress-specific QTL for SWU, suggesting that the control of transpiration was relevant to dry matter production under drought. We concluded that PEG-treated hydroponic culture is very effective for use in genetic analyses of drought tolerance at seedling stage.