花生FAD2基因家族表达分析及其对低温胁迫的响应

为了探究FAD2在花生低温响应中的作用,本研究从普通油酸花生中花16 (ZH16)和高油酸花生中花413 (ZH413)中克隆得到花生AhFAD2家族的全部基因,共7个。通过分析这些基因的表达模式发现,在ZH16和ZH413中各FAD2基因表达模式相似, AhFAD2-1A/B主要在花和发育的种子中表达, AhFAD2-3A/B主要在营养组织中表达, AhFAD2-4A/B主要在根和花中表达,表明AhFAD2基因在花生不同发育阶段和不同组织中发挥各自的生物学功能。在15℃下发芽6 d发现,ZH413的发芽率未显著下降,而ZH16的发芽率显著下降。种子萌发过程中,AhFAD2-1A/B和AhFAD2-4A/B均受低温诱导表达。在ZH16中AhFAD2-1A/B在低温诱导第6天开始显著上调表达,而在ZH413中第1天显著上调表达;在ZH16中AhFAD2-4A/B在低温诱导第3天出现显著上调表达,之后表达量下降,但在ZH413中第1天就显著上调表达,且始终维持在高水平表达。基于以上研究结果推测,高油酸花生在受到低温胁迫时,AhFAD2-1A/B编码蛋白失活,但AhFAD2-4A/B的高量表达在一定程度上弥补了这部分功能。同时也说明AhFAD2-1A/B功能的缺失并不是决定花生耐寒性的主要因素。本研究的开展为培育抗寒的高油酸花生品种奠定了理论基础,为高油酸花生在高纬度、高海拔地区推广提供了理论支持。     

Identification of quantitative trait loci and candidate genes controlling the tocopherol synthesis pathway in soybean (Glycine max)

A recombinant inbred line (RIL) population was used to identify quantitative trait loci (QTLs) and their candidate genes controlling the tocopherol (Toc) synthesis pathway. The RIL population was cultivated in field conditions in 3 years. A genetic map constructed using 1624 DNA markers was used for QTL analysis. We identified 22 QTLs for seed tocopherol contents and their ratios, of which two QTL clusters on chromosomes (Chr) 9 and 14 exerted consistent large effects on tocopherol composition across the 3 years. The QTL cluster localized on Chr 9 might correspond to γ-TMT3, which controls the conversion of γ-Toc into α-Toc. The QTL cluster localized on Chr 14 was novel, which might regulate the conversion of MPBQ (a precursor of δ-Toc) into DMPBQ (the precursor of γ-Toc). The effect of the QTL cluster on Chr 14 was validated in a pair of near isogenic lines, and its candidate gene was mined. The identified QTLs and their candidate genes might be used in breeding programmes to improve α-Toc content in soybean seeds.     

Genetic dissection of vitamin E content in rice (Oryza sativa) grains using recombinant inbred lines derived from a cross between 'Zhenshan97B' and 'Nanyangzhan'

Vitamin E (VE) is an important nutritional trait in rice grains. In order to dissect the genetic basis underlying VE content, a recombinant inbred lines population derived from 'Zhenshan 97B' and 'Nanyangzhan' was used for quantitative trait locus (QTL) analysis. Totally, 29 QTLs for six VE traits were identified in 2 consecutive years. Among those, five QTLs repeatedly detected in two years formed a cluster on chromosome 2, which was responsible for all five VE isomers. OsγTMT, the gene encoding γ‐tocopherol methyltransferase in rice, was located to the same region and treated as the candidate gene. Sequence analysis of alleles from two parents revealed many polymorphisms, including 19 single nucleotide polymorphisms (SNPs) and two insert/deletions (Indels) in the promoter region, two nonsynonymous SNPs in exons, and 25 SNPs and an Indel in introns. Besides, a QTL for δ‐tocotrienol and two QTLs for α‐tocopherol were repeatedly detected on chromosome 5 and 8, respectively, all three regions carrying no homologous genes involved in VE biosynthesis. These results could be useful in development of rice lines displaying desirable VE content.     

Influence of rare alleles of β‐carotene hydroxylase and lycopene epsilon cyclase genes on accumulation of provitamin A carotenoids in maize kernels

Vitamin A deficiency is one of the major health problems worldwide. Traditional yellow maize possesses very low provitamin A (proA) concentration in endosperm. The influence of rare alleles of β‐carotene hydroxylase (crtRB1) and lycopene epsilon cyclase (lcyE) genes capable of enhancing proA concentration was studied in four BC₂F₂ populations generated using subtropical inbreds and CIMMYT‐HarvestPlus lines. The occurrence of severe segregation distortion for crtRB1 gene was observed, while lcyE gene was segregated as per Mendelian ratio. Genotype with favourable allele of crtRB1 (CC) had a significant effect on β‐carotene (BC) (7.9‐fold), β‐cryptoxanthin (BCX) (twofold) and proA (5.5‐fold) accumulation, compared to unfavourable genotype (C⁺C⁺). Genotype with favourable allele of lcyE (LL) showed 2.1‐fold, 1.6‐fold and twofold significant enhancement in BC, BCX and proA, respectively, over unfavourable genotype (L⁺L⁺) in pooled analysis. Of the nine genotypes, double homozygote (CC/LL) had the highest mean BC (12.60 μg/g), BCX (4.44 μg/g) and proA (14.82 μg/g), and combined effect was significantly better than individual gene effects or any other combinations. The information generated here would be useful in designing strategy for proA enrichment in subtropical maize.     

Genetic analysis of antixenosis resistance to the common cutworm (Spodoptera litura Fabricius) and its relationship with pubescence characteristics in soybean (Glycine max (L.) Merr.)

The common cutworm (CCW, Spodoptera litura Fabricius) is one of the most serious pests of soybean (Glycine max (L.) Merr.). Previously, two quantitative trait loci (QTLs) for antibiosis resistance to CCW, CCW-1 and CCW-2, were detected in the resistant cultivar Himeshirazu. In this study, we conducted an anti-xenosis bioassay using a recombinant inbred population derived from a cross between a susceptible cultivar Fukuyutaka and Himeshirazu to perform QTL analysis. Two QTLs for antixenosis resistance, qRslx1 and qRslx2, were identified on Chrs 7 and 12, and the resistant alleles of qRslx1 and qRslx2 were derived from Himeshirazu and Fukuyutaka, respectively. The position of qRslx1 is similar to that of CCW-1. We also analyzed pubescence characteristics because they have been reported to be associated with soybean insect resistance. Two QTLs for pubescence length (on Chrs 7 and 12) and two QTLs for pubescence density (on Chrs 1 and 12) were identified. The pubescence QTLs on Chrs 7 and 12 were located near qRslx1 and qRslx2, respectively. These results suggest that the antixenosis resistance could be controlled genetically by the identified QTLs and that the pubescence characteristics might contribute to the soybean antixenosis resistance to CCW.     

Growth response of 98 barley (Hordeum vulgare L.) genotypes to elevated CO2 and identification of related quantitative trait loci using genome‐wide association studies

Elevated CO₂ (eCO₂) concentrations can stimulate crop growth, but little is known about intraspecific variability in the response to eCO₂ and the underlying genetics in cereals. Field experiments over two years with 98 barley genotypes were conducted in open‐top chambers (OTCs) under ambient CO₂ (400 ppm) and eCO₂ (700 ppm) concentrations. At crop maturity, different fractions of aboveground biomass (AGB) were measured, and genome‐wide association studies (GWASs) were conducted to identify quantitative trait loci (QTL). Averaged across all genotypes, eCO₂ significantly enhanced AGB by 15%, while the increase in culm and ear biomass alone was not significant. The AGB response to eCO₂ of the individual genotypes ranged from c. ?36% to +95% compared with ambient CO₂ (aCO₂), showing a large variability of growth responses. In GWAS, 51 associations between SNP markers and the relative changes (eCO₂/aCO₂) in biomass were detected on different chromosomes. Loci potentially involved in biomass alterations under eCO₂ were identified. The wide range of variability in responses might be exploited by marker‐based breeding for climate‐resilient barley.     

Modelling approach for the quantitative variation of sex expression in monoecious hemp (Cannabis sativa L.)

Sex expression is of primary importance for the genetic improvement and production of monoecious hemp: masculinized phenotypes are associated with higher fungal sensitivity, and feminized phenotypes with higher seed yields. However, sex expression varies quantitatively among plants and nodes and with time. Here, we developed eight variables that characterize the sex expression in monoecious hemp to dissect its genetic determinism. The monoecy degree (MD), ranging from 1 (mostly male flowers) to 5 (mostly female flowers), was recorded for each node of 167 plants, at 6 times at 1‐week intervals. Two types of longitudinal variables were constructed: ‘synthesis’ (mean MD and percentages of nodes of each MD) and ‘structure’. The latter consisted of the parameters of a logistic curve describing MD as a function of the node position. An r‐square of 0.97 was obtained between the estimated and observed MD values, and the logistic parameters were weakly correlated with each other and with the synthesis variables. Therefore, we conclude that the present modelling approach is relevant for characterizing the sex expression in monoecious hemp.     

Carotenoid accumulations and carotenogenic gene expressions in the petals of Eustoma grandiflorum

Eustoma grandiflorum is one of the leading cut‐flowers in Japan. There are market demands for cultivars with deep‐yellow flowers, but they have never been bred successfully. By investigating the carotenoid accumulation and carotenogenic gene expressions, this study attempted to explore the reasons that block the formation of deep‐yellow colour in Eustoma. High performance liquid chromatography analysis showed that the carotenoid compositions in petals were similar to those in leaves, accumulating mainly lutein, violaxanthin and β‐carotene. The total carotenoid contents decreased as the petals matured in all the cultivars tested. Quantitative real‐time PCR analysis showed that the expression levels of PSY, LCYB, ZDS and LCYE showed significant differences between white and pale‐yellow petals or between petals and leaves, indicating that these enzymes may play a key role in the carotenoid biosynthesis in E. grandiflorum. The expression levels of CCD4 were high in both pale‐yellow and white petals during development, suggesting that carotenoid degradation activity is high in the petals. We then conclude that the total carotenoid accumulation level could be determined by the balance between carotenoid biosynthesis and degradation activities.     

Photosynthesis is Reduced,and Seeds Fail to Set and Fill at Similar Soil Water Contents in Grass Pea (Lathyrus sativus L.) Subjected to Terminal Drought

Grass pea (Lathyrus sativus L.) is an indeterminate grain legume considered adapted to dry environments, but the mechanisms of its adaptation are not well understood. Grass pea plants were exposed to terminal drought from podding, and the development of water deficit was measured together with its effects on leaf photosynthesis, stomatal conductance, carbon remobilisation to the seeds, flower production and abortion, pod production and abortion, seed set, seed growth and the neurotoxin β‐N‐oxalyl‐L‐a, β‐diaminopropionic acid (β‐ODAP) concentration. Predawn leaf water potential (Ψ_leaf), stomatal conductance (gs), rate of leaf photosynthesis (Pn), flower production, pod production, filled pod number, seed number, seed size and yield decreased, while flower abortion, pod abortion and seed abortion increased, and the concentration of β‐ODAP was unchanged under terminal drought conditions. gs and Pn began to decrease at a higher plant‐available soil water content (PAWC) (67.2 ± 2.3 % and 62.9 ± 2.3 %) than Ψ_leaf (43.7 ± 1.1 %). Flowers and pods ceased being produced only when the PAWC decreased below 40.1 ± 4.6 % and 35.3 ± 3.0 %, respectively, but seed set and seed growth ceased when PAWC decreased below 55.5 ± 1.6 % and 58.0 ± 3.7 %, respectively. The mobilization of ¹³C labelled assimilates from the stems was greater under terminal drought than under well‐watered conditions, but the transfer to the seed was small. We conclude that seed set and seed growth decreased as the soil dried due to a reduction in current photosynthesis as a result of stomatal closure.     

Quantitative trait loci for quality and agronomic traits in two advanced backcross populations in oat (Avena sativa L.)

Using the advanced backcross quantitative trait loci (AB‐QTL) strategy, we successfully transferred and mapped valuable allelic variants from the high β‐glucan (BG) accession IAH611 (PI 502955), into the genome of cultivar ‘Iltis’. By backcrossing one BC₁F₁ plant to ‘Iltis’, we developed two BC₂F_2‐6 populations A and B, comprising 98 and 72 F₂‐individuals, respectively. Genotyping of BC₂F₂ individuals with predominantly AFLP markers resulted in 12 linkage groups with a map size of 455.4 cM for Population A and 11 linkage groups with a map size of 313.5 cM for Population B. Both populations were grown at three sites in Germany over a three‐year period. Individuals were then phenotyped for 13 traits including grain yield (YD) and β‐glucan content (BG). QTL analysis via stepwise regression detected a total of 33 QTLs, most of which were clustered in three linkage groups. Two dense linkage groups A1 and B13 were found to be putatively homologous to groups KO_6 and KO_11 of the ‘Kanota’/‘Ogle’ map, respectively.     

元麦β-葡聚糖含量的差异性及其影响因素的研究进展

元麦膳食纤维含量丰富,β-葡聚糖含量高,保健功效强,是重要的功能性农产品。为了全面了解不同地区元麦β-葡聚糖含量的差异性及其影响因素,本研究对全国7个不同省份元麦棱型、粒色、β-葡聚糖含量进行了整理和归纳,分析了气候、遗传育种和栽培措施等方面对元麦β-葡聚糖含量的影响。针对有关元麦β-葡聚糖合成筛选的有效基因少,气候与栽培措施对元麦β-葡聚糖的影响较大等问题,提出如下解决建议：（1）通过全基因组关联分析定位筛选相关位点与候选基因,挖掘与利用和β-葡聚糖合成相关的关键或主导基因,为元麦营养育种提供分子基础。（2）实际生产中根据当地气候条件,选择高产高β-葡聚糖含量的加工用营养元麦品种,采用适宜的栽培措施,缓解不良生长环境的影响。元麦分子育种和适宜的栽培措施有助于提高元麦生长适应能力,稳定元麦β-葡聚糖的含量,保障高品质元麦原料,推动健康农业的发展。     

Contribution of the grain size QTL GS3 to yield properties and physiological nitrogen-use efficiency in the large-grain rice cultivar ‘Akita 63’

The development of crop varieties with high nitrogen-use efficiency (NUE) is thought to be important in achieving sustainable cereal crop production. The high yield large-grain rice cultivar Oryza sativa L. ‘Akita 63’ (temperate japonica) has high physiological NUE (PNUE) for grain yield (GY). Our previous study revealed that a large-grain allele of GS3 is present in ‘Akita 63’. Here, we verified the influence of GS3 on the yield properties and PNUE for GY in ‘Akita 63’. The frequency distribution of brown rice length in F₂ crosses of ‘Iwate 75’ and ‘Akita 63’ showed a continuous distribution that could be explained by GS3. A near-isogenic line was developed to substitute the GS3 segment of ‘Koshihikari’, which harbours a normal-sized grain allele, in the genetic background of ‘Akita 63’ and the line was designated as Akita63NILGS3-Koshihikari. Compared with Akita63NILGS3-Koshihikari, ‘Akita 63’ exhibited a significantly increased grain length, single brown grain weight and GY, although no significant differences were observed in the nitrogen content and above-ground biomass per unit of cultivated area. These results indicate that the GS3 large-grain allele is a contributing factor to high PNUE for GY in ‘Akita 63’. These findings will facilitate the development of nitrogen-efficient rice varieties.     

Alleles at a quantitative trait locus for stem solidness in wheat affect temporal patterns of pith expression and level of resistance to the wheat stem sawfly

The stem solidness trait in wheat has been the most effective mechanism for management of the wheat stem sawfly (WSS) for six decades. However, recent results have shown that in certain genotypes, the degree of stem solidness is not a useful indicator of WSS resistance. A morphological characterization of solidness expression indicated that in the genotype ‘Conan’, very solid pith undergoes rapid retraction during stem maturation, resulting in significantly less solidness at maturity. In other solid‐stemmed genotypes, including the standard WSS‐resistant cultivar ‘Choteau’, dense pith in the stem remains nearly unchanged throughout plant development. In cage trials, ‘Conan’ plants were less preferred for oviposition by the WSS when paired with ‘Choteau’ plants. Field bioassays using near‐isogenic lines differing for alleles at Qss.msub‐3BL showed that the Conan allele provides higher levels of early stem solidness and rapid pith retraction during stem maturation. These results suggest that the traditional approach for increasing WSS resistance by selecting for increasing stem solidness needs to be modified to consider temporal variations in pith expression associated with alleles at Qss.msub‐3BL.     

Temperature and resistance gene interactions in the expression of resistance to Blumeria Graminis f. sp. Tritici

The influence of temperature on the resistance to powdery mildew, caused by Blumeria graminis (DC.) E.O. Speer f. sp. tritici Em. Marchal, was evaluated on six wheat cultivars/lines, Axminster/8^* Chancellor (Cc) (Pm1), Chul/8^*Cc (Pm3b), Yuma/8^*Cc (Pm4a), VPM1 (Pm4b), Kavkaz (Pm8), and Chancellor, at two temperatures of 25EC and 15EC with two isolates of powdery mildew. Resistance, based on the components of latency period, infection type, colony number per unit leaf area, per cent infected leaf area, and AUDPC, was much less effective at 15EC than at 25EC for VPM1 and Yuma/8^*Cc, but no resistance shift was observed for Kavkaz and Axminster/8^*Cc, indicating the difference of temperature-sensitivity among Pm genes or among the interactions of individual host genes with corresponding powdery mildew pathogen genes. This revised version was published online in July 2006 with corrections to the Cover Date.