大豆蛋白质有关性状的QTL定位

以科丰1号×南农1138-2组合衍生的184个重组自交家系(简称RIKY)和(Essex×ZDD2315)×ZDD2315衍生的114个BC₁F₂家系(简称BIEX)为材料,对蛋白质含量、蛋油总量与油脂含量,11S、7S、11S/7S,11S-1~11S-4, 7S-1~7S-6等4组16个性状利用WinQTL Cartographer Ver.2.5的复合区间作图法(CIM)、多区间作图法(MIM)和IciMapping Ver.2.0的完备区间作图法(ICIM)进行QTL分析, 结果表明：(1) 在RIKY和BIEX群体分别定位到17⁺个和21⁺个QTL,合计38⁺个QTL;在RIKY有蛋白、油脂、蛋油总量QTL11个,在11S和7S亚基组上分别只有1⁺和3⁺个;在BIEX有前性状QTL2⁺个,有后性状QTL分别9⁺和6⁺个;(2) 两群体16个性状上均没有检测到共享的QTL,说明两群体的蛋白质有关性状具有完全不同的遗传基础;RIKY的两个亲本间蛋白、油脂和蛋油总量有明显遗传差异,但在亚基组上遗传差异不大,而BIEX则反之;(3) 4组总、分性状中,两群体一致表现出蛋白、油脂和蛋油总量和11S、7S和11S/7S比值两组在总、分性状间共享QTL(共同遗传基础), 而11S亚基组和7S亚基组两组性状在总、分性状间无共享的QTL;(4) 蛋白质有关性状QTL定位结果和分离分析结果共同表明这类性状主效基因和微效基因均占较大比重,要考虑两者兼用的育种方法。     

大豆蛋白质有关性状遗传的分离分析

采用科丰1号×南农1138-2衍生的重组自交家系群体(RIKY)和(Essex×兴县灰布支)×兴县灰布支回交自交衍生群体(BIEX)为材料,应用SDS-PAGE电泳测定11S、7S、11S/7S比值及其10个亚基组的相对含量,用凯氏(Kjeltec)自动定氮仪测定蛋白质含量、自动索氏(Soxtec)抽提仪测定油脂含量。结果表明: (1) 两群体蛋白质组分及其亚基组有关性状以及油脂与蛋油总量等均有不同程度超亲分离,双亲间存在不同程度的位点互补。(2) 蛋白质含量遗传,两群体均为1对主基因+多基因模型,主基因和多基因遗传率分别为31.3%~40.9%和37.2%~53.7%。蛋油总量均为3对主基因+多基因模型,主、多基因遗传率分别为59.9%~66.6%和23.2%~27.9%。油脂含量为2~3对主基因+多基因模型,两类遗传率分别为48.6%~71.7%和4.2%~29.7%。(3) 11S组分均为3对主基因+多基因模型,两类遗传率分别为14.3%~60.7%和17.0%~50.7%。7S组分均为3对主基因+多基因模型,两类遗传率分别为34.5%~44.1%和21.5%~45.1%。11S/7S比值遗传均为2对主基因+多基因模型,两类遗传率分别为56.6%~74.8%和10.1%~20.1%。(4) 11S的4个亚基组依次分别为2~3对主基因+多基因、2对主基因+多基因、2对主基因+多基因、2~3对主基因+多基因模型。(5) 7S的6个亚基组依次分别为1~2对主基因+多基因、2对主基因+多基因、2~3对主基因+多基因、3对主基因+多基因、1~2对主基因+多基因和2对主基因+多基因模型。所有16个性状都由主基因和多基因控制,其中有10个性状两群体具相同的主基因数,其他6个性状两群体间相差1对主基因,两群体间遗传模型大同小异。这些相关性状的育种既要利用主基因还必须利用微效多基因,要考虑兼用两者的育种方法。     

大豆胁迫相关蛋白GmSAP3基因的克隆和表达分析

胁迫是影响植物生长发育和农作物产量的重要因素.胁迫相关蛋白(stress associated protein,SAP)是一类锌指蛋白,广泛参与植物发育和胁迫响应.为了研究大豆SAP基因的功能,本研究以'商豆1201'为材料克隆了GmSAP3基因,并进行了生物信息学分析,采用RT-PCR分析该基因在大豆不同组织和温度胁迫下的表达模式.结果 表明,GmSAP3基因CDS序列全长为513 bp,编码170个氨基酸,GmSAP3蛋白等电点pI为6.79,分子量为18.30568kD;序列分析发现,GmSAP3含有2个保守结构域(zf-A20和ZnF-AN1),序列比对和进化分析发现GmSAP3与GmSA P26亲缘关系最为接近.RT-PCR结果表明,GmSAP3在大豆不同组织中均有表达,其中根和叶片表达较高;GmSAP3受高温胁迫诱导表达,推测该基因在大豆胁迫响应中具有重要作用.本研究为GmSAP3基因功能研究提供了一定的理论支持.     

大豆百粒重QTL定位

大豆百粒重是产量构成的重要因素之一,与产量呈正相关。本研究以溧水中子黄豆和南农493-1的504个F2正反交单株及其亲本间具有多态性的150个SSR标记信息构建连锁图谱,2008年分别在江苏南京和山东临沂两地种植其衍生的正反交F2:4家系,鉴定其百粒重,应用Win QTL CartographerV2.5复合区间作图法和两地正反交联合分析进行QTL定位。结果表明,复合区间作图法检测到16个主效QTL,联合分析检测到24个主效QTL、环境效应与细胞质效应、1个环境×QTL互作和12个细胞质×QTL互作。其中,两方法共同检测到10个主效QTL,正反交群体在两地中共同检测到3个主效QTL;Meta分析发现与其他研究一致的4个QTL。这些结果为大豆产量遗传与标记辅助育种实践提供理论基础。     

紫外线-B辐射增强对大豆生长、发育、色素和产量的影响

在田间条件下，模拟20%平流层臭氧层的减薄，研究了增强紫外线-B辐射(280～320nm)(UV-B)对10个大豆品种生长、发育、色素和产量的影响。结果表明：(1)增强的UV-B辐射使一些大豆品种的物候期发生改变。(2)明显降低多数品种光合作用色素和类胡萝卜素的含量，增加所有品种类黄酮类化合物的含量。(3)大豆品种的株高、根茎叶生物量     

大豆四向重组自交系群体蛋白质含量与油分含量QTL定位

蛋白质和油分含量是大豆重要的育种目标,蛋白质和油分含量QTL定位和优异等位变异的发掘对大豆分子设计育种具有重要意义。本研究以(垦丰14×垦丰15)×(黑农48×垦丰19)衍生的后代株系为材料,构建含有204个株系的大豆四向重组自交系群体,利用区间作图法,应用前期构建的SSR遗传图谱,对2013、2014和2015年在哈尔滨和克山2地共8个环境下的蛋白质和油分含量进行QTL定位分析。结果表明,8个环境中检测到29个蛋白质含量QTL和39个油分含量QTL。在所定位的蛋白质含量QTL中,有5个能够在2个以上环境被定位到,这些蛋白质含量QTL分布在 A1、D2、J、N和O等6个连锁群上,对表型效应的贡献率为 7.65%~20.08%,其中qPC-A1-1、qPC-D2-1、qPC-J-1和qPC-O-2的贡献率在10%以上。在39个油分含量QTL中,有10个在多环境下被重复检测到,这些QTL分布在8个(A1、A2、B1、D1b、G、I、J、N)连锁群上,对表型效应的贡献率为7.30%~25.68%,其中qOC-A2-1、qOC-B1-1、qOC-G-1和qOC-J-1的贡献率在10%以上。     

Deficit Irrigation of Soya Bean [Glycine max (L.) Merr.] in a Sub-humid Climate

An experiment was conducted to investigate the influence of different levels of water deficit on yield and crop water requirement of soya beans in a sub‐humid environment (Southern Marmara region, Bursa, Turkey) in 2005 and 2006. One full‐irrigated treatment (T₁), one non‐irrigated treatment (T₅) and three different deficit irrigation (T₂ = 25 % water deficit, T₃ = 50 % water deficit, T₄ = 75 % water deficit) treatments were applied to ‘Nova’ soya bean planted on a clay soil. Non‐irrigated and all deficit irrigation treatments significantly reduced biomass and seed yield and yield components. The full‐irrigated (T₁) treatment had the highest yield (3760 kg ha^?1), while the non‐irrigated (T₅) treatment had the lowest yield (2069 kg ha^?1), a 45.0 % seed yield reduction. T₂, T₃ and T₄ deficit irrigation treatments produced 11.7–27.4 % less seed yield than the T₁ treatment. Harvest index showed less and irregular variation among irrigation treatments. Both leaf area per plant and leaf area index were significantly reduced at all growth stages as amount of irrigation water was decreased. Evapotranspiration increased with increased amounts of irrigation water supplied. Our results indicate that higher amounts of irrigation resulted in higher seed yield, whereas water use efficiency and irrigation water use efficiency values decreased when irrigation amount increased.     

The Interaction of Sowing Date and Water Availability in Determining Plant Architecture, Fruiting Pattern and Yield Components of Soybean (Glycine max L. Merr.)

A 2-year trial has been carried out in northern Italy on soybean (cv. Hodgson) grown in lysimeters, comparing three soil water regimes (well-watered conditions and water stress during vegetative and reproductive stages) at two sowing dates. Plant evapotranspiration and water uptake depth were calculated from volumes of water independently supplied to eight lysimeter layers; at harvest, plant architecture, yield components and fruit distribution along the main stem and lateral branches were evaluated.
Although water stress intensity was not severe, crop evapotranspiration and water uptake depth were severely restricted by water shortage. Both low water-availability and late sowing significantly modified the architecture of plants, decreasing total height, number and length of internodes and lateral branches. Seed allocation along the stem was shifted downwards both by delaying the sowing date and by reducing the water supply; the component most responsible for yield decrease was the number of pods per plant, while unit seed weight was only slightly affected by water stress. Grain yield reduction was higher when water availability was inadequate during the reproductive phase in the early-sown crop and during the vegetative stage in the late-sown crop. This suggests that the intensity of the water shortage, plant phenological stage of stress application, as well as the date of stress application within the growing season determine the yield response of soybean.     

Mineral Nitrogen Availability and Isoflavonoid Accumulation in the Root Systems of Soybean (Glycine max (L.) Merr.)

Isoflavonoids, as plant-to-bacteria signal molecules, play an important role in the establishment of the soybean ( Glycine max (L.) Merr.)– Bradyrhizobium japonicum nitrogen (N) fixing symbiosis. They are essential to the development of effective root nodules and responsible for inducing the nod genes of B. japonicum . Because N affects a broad range of infection events, especially the symbiotic events occurring within 18 h of inoculation, it is reasonable to hypothesize that mineral N disrupts the interorganismal signal exchange between soybean host plants and B. japonicum . High performance liquid chromatographic (HPLC) analysis of root extracts of soybean, inoculated with B. japonicum or not, grown with various levels of mineral N in the rooting medium were performed to test this hypothesis. The results of these studies indicated that: (1) at early plant growth stages (before the onset of N fixation), a strong negative relationship between N application and soybean root isoflavonoid (genistein and daidzein) concentrations existed; (2) although isoflavonoid (genistein and daidzein) concentrations in both inoculated and non-inoculated soybean root systems were generally decreased by N application, at very low N levels (10 mg N l⁻¹) genistein in the non-inoculated plant roots was not decreased relative to the 0 N plants; (3) averaged over all mineral N treatment levels and sampling times, inoculation of soybean with B. japonicum increased root daidzein concentrations (P > 0.05), but did not affect genistein. Overall, N application reduced the isoflavonoid concentration of soybean root systems, which probably plays a part in the regulation of soybean nodule formation by available N.     

大豆细胞质雄性不育系MADS-box基因的分离分析

采用cDNA-AFLP差异显示技术对大豆细胞质雄性不育系NJCMS2A与其保持系NJCMS2B间基因差异表达进行研究,结果从NJCMS2A花蕾中分离到一个差异表达片段,对该差异片段进行克隆、测序和序列比对分析,Blast检索结果显示它与大豆基因组中Gm13上g29510.1 cDNA片段的同源性达98.7%,与大豆中一个MADS-box基因的同源性达98%,氨基酸序列比对结果表明它与大豆中一个MADS-box蛋白有96%的同源性,与豌豆中MADS-box M7蛋白有83%的同源性,与苦瓜中MADS-box2蛋白有88%的同源性,与海岛棉典型的MADS-box基因编码的AGAMOUS蛋白保守区有83%的同源性,进一步对其氨基酸序列进行结构和功能预测显示该差异片段具有MADS-box转录因子的典型结构域K-box,证明其编码蛋白为一MADS-box转录因子,半定量RT-PCR分析结果显示其在NJCMS2A花蕾中表达量很高,而在NJCMS2B花蕾中表达量很低,推测该差异片段可能与大豆细胞质雄性不育有关。     

Genotype x Bradyrhizobium japonicum strain interactions in dinitrogen fixation and agronomic traits of soybean (Glycine max L. Merr.)

Summary Bradyrhizobium japonicum strain G49 has been the only inoculum used in French soils. Soybean (Glycine max L. Merr.) cultivars were selected and tested according to their performances with this rhizobial strain. The aim of the present study was to determine the consequences of strain substitution on N₂ fixation abilities of various genotypes. Three genotypes and cultivar Weber, in combination with B. japonicum strain G49 or SMGS1, were cultivated in pots and tested for nitrogenase activity under differing nitrogen nutrition conditions. The reliability of ARA (acetylene reduction activity) measurement for assessing symbiotic nitrogen fixation under the experimental conditions used was checked. Genotypic variability for symbiotic fixation activity was observed with each strain under soil culture conditions; important genotype x strain interactions were also involved. These results were corroborated for the protein yield and other yield component performances of the various genotype-strain associations. Thus, in France, the replacement of strain G49 with another one might result in the alteration of the relative agronomic performances of the soybean cultivars, since N₂ fixation is considered as a major factor of soybean productivity.     

植酸酶phyA基因的密码子优化及其在大豆中的表达

植酸是植物源食品中的主要抗营养成分, 降低植酸含量可有效提高大豆的营养利用率。本文根据大豆密码子使用偏好性, 对无花果曲霉植酸酶phyA基因进行密码子优化, 人工合成了适合在大豆中表达的phyA(b)基因。以pCAMBIA3301为骨架, 构建由大豆凝集素基因启动子和信号肽序列调控的植物表达载体pCBPS-phyA(b)。用农杆菌介导法遗传转化吉林35大豆子叶节。PCR检测表明目的基因已初步整合至大豆基因组中; bar试纸条表明所有阳性植株中均能检测到bar基因的蛋白产物; 除草剂叶片涂抹显示野生型的叶片出现黄化或枯萎现象, 而转基因植株叶片表现正常, 具除草剂抗性; 以半定量RT-PCR共筛选到13株转phyA和19株转phyA(b)阳性转基因大豆植株。通过对转基因大豆T3种子中植酸酶活性、无机磷和植酸磷含量等检测, 证明人工基因phyA(b)比phyA在大豆种子中所表达的植酸酶具有更高的活性, 说明密码子优化有利于提高外源基因的表达。     

大豆光周期反应与生育期基因研究进展

20世纪20年代,植物学家Garner与Allard在研究大豆与烟草等植物的光反应时发现了植物光周期现象。大豆作为模式植物对光周期现象的理论形成起了重要作用。但大豆基因组的复杂性及与相关功能基因关系的不明确性严重阻碍了学者对大豆光周期现象本质的认识。近年来,随着控制大豆生育期主要QTL基因的相继克隆,特别是对大豆生育期贡献最大的E1基因的成功破译,学者们逐步认识到大豆光周期调控开花的独特性。遗传学及分子生物学研究表明,大豆中具有拮抗关系的E1和FT基因位于大豆光周期调控开花主要通路的中心节点(integrator),但两者间的作用机制及相关的调节因子尚待明晰。对大豆光周期反应及生育期基因的深入研究,在生产实践上可为大豆品种的栽培区划、合理布局及分子育种等提供理论依据。     

Soybean (Glycine max (L.) Merr.) growth and development response to CO2 enrichment under different temperature regimes

The carbon dioxide (CO₂) concentration of the global atmosphere has increased during the last decades. This increase is expected to impact the diurnal variation in temperature as well as the occurrence of extreme temperatures. This potentially could affect crop production through changes in growth and development that will ultimately impact yield. The objective of this study was to evaluate the effect of CO₂ and its interaction with temperature on growth and development of soybean (Glycine max (L.) Merr., cv. Stonewall). The experiment was conducted in controlled environment chambers at the Georgia Envirotron under three different temperatures and two CO₂ regimes. The day/night air temperatures were maintained at 20/15, 25/20 and 30/25 °C, while the CO₂ levels were maintained at 400 and 700 ppm, resulting in six different treatments. Plants were grown under a constant irradiance of 850 μmoles m⁻² s⁻¹ and a day length of 12 h; a non-limiting supply of water and mineral nutrients were provided. Five growth analyses were conducted at the critical development stages V4, R3, R5, R6 and R8. No differences in start of flowering were observed as a function of the CO₂ level, except for the temperature regime 25/20 °C, where flowering for the elevated CO₂ level occurred 2 days earlier than for the ambient CO₂ level. For aboveground biomass, an increase in the CO₂ level caused a more vigorous growth at lower temperatures. An increase in temperature also decreased seed weight, mainly due to a reduction in seed size. For all temperature combinations, final seed weight was higher for the elevated CO₂ level. This study showed that controlled environment chambers can be excellent facilities for conducting a detailed growth analysis to study the impact on the interactive effect of changes in temperature and CO₂ on soybean growth and final yield.     

Soya Bean (Glycine max L. Merr.) Genotype Response to Early-season Flooding: II. Aboveground Growth and Biomass

Soya bean is often grown in regions prone to periodic flooding, thus selecting cultivars that maintain production under waterlogged conditions is desirable. An experiment involving flooded soya beans was planted in southern Florida to examine (1) stem and leaf growth; (2) morphological adaptations; and (3) the relationship between early‐season and late‐season flood tolerance in flooded soya beans. Eleven soya bean genotypes previously defined as tolerant or sensitive to flooding were subjected to three treatments at 21 days after sowing (DAS): (1) no flood, (2) 2‐week flood and (3) 4‐week flood. All plants were harvested 49 DAS. Flooded plants exhibited lower stem dry weights but greater partitioning to the stem. Non‐flood treatments had greater leaf dry weight, leaf area and partitioning to leaves than flooded plants. There were positive correlations of genotype stem dry weight and leaf dry weight to early‐season flood tolerance but stem partitioning was negatively correlated with early‐season flood tolerance. Genotypic rankings of early‐season flood tolerance in this study were not correlated with earlier studies basing flood tolerance on seed yield. Our study highlights the range of soya bean morphological adaptations in response to flood. However, our results indicate that early‐season screening may not be an accurate predictor of soya bean genotypic response to late‐season flood.     

Response of Soybean [Glycine max (L.) Merr.] Cultivars to Genistein-Preincubated Bradyrhizobium japonicum: Nodulation and Dry Matter Accumulation under Canadian Short-Season Conditions

The addition of genistein, a plant-to-bacteria signal molecule, to Bradyrhizobium japonicum cells prior to use as inocula has been shown to increase nodule number and promote soybean N₂ fixation at low root zone temperatures. Previous greenhouse and field experiments involving only two cultivars have indicated that soybean [ Glycine max (L.) Merr.] cultivars can vary in their response to genistein application. The objective of this study was to evaluate a range of soybean cultivars for response to genistein application under short-season cool-spring Canadian conditions. A 2-year field study was conducted in 1997 and 1998 with a range of soybean cultivars recommended for Quebec. The 11 cultivars tested represented a range of yield potentials and maturity groups. They were inoculated with genistein-preincubated B. japonicum inocula or regular inocula, applied into the furrow at the time of planting. The results of these experiments indicated that neither maturity nor yield was correlated with increases in nodulation, biomass, and plant total nitrogen content resulting from genistein treatment and that all maturity groups responded to genistein application in essentially the same way. Thus, response of soybean cultivars to genistein addition is regulated by genotype characteristics other than maturity or yield level.     

Growth and Development of Soybean (Glycine max [L.] Merr.) Cultivars under Shade in Coconut Garden

The growth and development of seven soybean cultivars were studied under shade in a coconut garden. Higher shoot height, internodal elongation and lower leaf area index were the most significant growth changes noticed under shade. Leaf net photosynthesis, crop growth rate and seed yield were also reduced under shade. The cultivars CO 1, UGM 30 and UGM 37 recorded higher yield under shade when compared to other cultivars tested.     

SSR marker tightly linked to the Ti locus in Soybean [Glycine max (L.) Merr.]

Soybean is a major source of protein meal in the world. Soybean kunitz trypsin inhibitor (SKTI) protein is a responsible for the inferior nutritional quality of unheated or incompletely heated soybean meal. The primary objective of this research was to identify DNA markers linked to the Ti locus controlling presence and absence of kunitz trypsin inhibitor protein. Two mapping populations were developed. Population 1 was derived from a cross between cultivar Jinpumkong2 (TiTi) and C242 (titi). Population 2 was made from a mating between cultivar Clark (TiTi) and C242. The F₁ plants were grown in the greenhouse to produce F₂ seeds. Each F₂ seed from F₁ plants was analyzed electrophoretically to determine the presence of the SKTI protein band. One-thousand RAPD primers, 342 AFLP primer sets, and 35 SSR primers were used to map Ti locus in population 1 and 2. The presence of SKTI protein was dominant to the lack of a SKTI protein and kunitz trypsin inhibit protein band was controlled by a single locus. Twelve DNA markers (4 RAPD, 4 AFLP, and 3 SSR) and Ti locus were found to be genetically linked in population 1 consisted with 94 F₂ individual plants. Three SSR markers (Satt409, Satt228, and Satt429) were linked with Ti locus within 10 cM. Satt228 marker was tightly linked with Ti locus. Satt228 marker was tightly linked within 0–3.7 cM of the Ti locus and may be useful in a marker assisted selection program.     

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.     

Effect of Seed Coating on Yield Attributes and Yield of Soybean (Glycine max L.)

Investigations were carried out during March to June 1991 to evaluate the effect of seed coating with organic, inorganic nutrients and biofertilizers on yield attributes and yield of soybean under three environments with different soil types viz., clay loam, sandy clay loam and sandy loam. Seed coating with bio-digested slurry 50 per cent, superphosphate 2 per cent, Bradyrhizobium 2 per cent, and Phosphobacteria 2 per cent (w/w of seed) increased the number of filled pods and grain yield by 29.6 and 37.2 per cent, respectively over the uncoated. There was a significant increase in the test weight of grains due to seed coating with Phosphobacteria. Environment with clay loam soil was found to be highly responsive to seed coatings.