南方大豆品种炭疽病抗性鉴定及抗病相关基因表达分析

为明确南方大豆品种对炭疽病抗性的差异,并分析其抗病相关基因的表达规律,在温室内种植华南农业大学近年来育成和收集到的100份南方春大豆品种,苗期喷洒平头炭疽菌孢子悬浮液,根据叶片病斑面积计算病斑率并划分为6个抗病级别。利用qRT-PCR技术检测典型抗病和感病品种接种平头炭疽菌不同时间后,病程相关蛋白基因、水杨酸途径基因、茉莉酸/乙烯途径基因和多聚半乳糖醛酸酶抑制蛋白基因等抗病相关基因的相对表达量变化情况。结果表明:共鉴定到6份抗病品种、32份中抗品种、57份中感品种和5份感病品种。抗性品种桂1701和感病品种横线早熟豆被平头炭疽菌侵染后抗病基因的表达规律存在很大差异。GmPR2、GmPR4、GmNPR1、GmPAL2-1和GmPAL4在抗病品种的表达量整体较高,GmPR3、GmPR12、GmPAL1-1和GmPAL3-1在感病品种中的表达量整体较高。GmPAL2-1和GmPAL3-1基因在抗病品种中的相对表达量分别在接种后12和6 h极显著升高,而在感病品种中则分别在接种后24和48 h才显著升高。GmPAL2-3在抗病品种中不表达,而在感病品种中表达;GmPGIP2在抗病品种中表达,而在感病品种中不表达。抗病品种中GmLOX7和GmLOX8基因的表达量是感病品种中的400~1 000倍。接种平头炭疽菌后48 h,GmPR2、GmPR4、GmPR5、GmPR10、GmNPR1、GmNAC4和GmPGIP3基因在抗病品种中的相对表达量显著升高,而在感病品种中显著降低。结果显示,桂1701、桂夏1号、南夏豆25、桂夏3号、华夏5和天长小青豆为抗炭疽病大豆品种。不同大豆品种受到平头炭疽菌侵染后,抗感品种间抗病相关基因的表达情况存在较大差异,抗病品种对该病菌侵染的反应更快,推测大豆炭疽菌侵染后48 h是大豆抗病基因响应侵染的关键时间点。

纳米氧化锌缓解大豆铝胁迫生理机制

【目的】探究在铝(Al)胁迫下不同含量的纳米氧化锌(Zinc oxide nanoparticles,ZnO NPs)对大豆Glycine max(Linn.)Merr.生理特性的影响,为金属纳米材料在农业上的应用提供一定参考。【方法】采用盆栽试验,选择耐Al品种‘华春2号’和普通品种‘华春6号’,在0.3 g/kg Al胁迫处理下,施加不同剂量的ZnO NPs(0、25、50、100和150 mg/kg),探究ZnO NPs对大豆生理指标(鲜质量、根长和叶绿素含量)、总超氧化物歧化酶(Total superoxide dismutase,T-SOD)活性和丙二醛(Malondialdehyde,MDA)浓度的影响。【结果】Al胁迫显著降低了‘华春6号’的鲜质量和根长,显著增加了其MDA浓度。对于‘华春2号’,Al胁迫显著增加了其叶绿素a和叶绿素b含量,而对其他指标无明显影响。无Al胁迫条件下施用ZnO NPs,均提高了‘华春6号’和‘华春2号’的鲜质量、根长和T-SOD活性。而在Al胁迫下施加不同剂量的ZnO NPs使‘华春6号’的鲜质量和根长分别增加13.2%~100.4%和7.8%~35.8%,而‘华春2号’的鲜质量在150 mg/kg ZnO NPs处理下达到最高值。在25 mg/kg ZnO NPs处理下,‘华春6号’和‘华春2号’的叶绿素a含量均达到最高值。不同含量的ZnO NPs对‘华春6号’的叶绿素b含量无显著影响,而显著降低了‘华春2号’的叶绿素b含量,在100 mg/kg ZnO NPs下达到最低值(3.5 mg/g)。‘华春6号’的T-SOD活性随ZnO NPs含量的增加而增加,‘华春2号’的T-SOD活性在50 mg/kg ZnO NPs时达到峰值(820 U/g),之后随ZnO NPs含量的增加出现下降的趋势。‘华春6号’和‘华春2号’的M DA浓度最小值分别出现在25和50 mg/kg ZnO NPs处理。【结论】Al胁迫严重影响大豆的生长发育,施用ZnO NPs可以在一定程度上缓解Al胁迫对大豆植株产生的负面作用,改善植株的生长发育。

大豆GmGST7基因耐酸铝功能研究

【目的】耐酸铝基因GsMYB7过表达转化大豆品种‘华春6号’后,从转基因株系的表达谱中获得目标基因GmGST7,该基因受酸铝胁迫诱导上调,且位于GsMYB7基因下游,进一步分析其耐酸铝功能,以期提高大豆酸铝耐受能力。【方法】采用生物信息学方法分析GmGST7基因的碱基序列、蛋白结构域和构建系统进化树。通过烟草叶片瞬时转化法完成亚细胞定位。通过RT-qPCR分析该基因组织表达特异性。设计0、25、50、75和100μmol/L 5个AlCl_(3)浓度梯度,研究GmGST7对酸铝胁迫的响应。在50μmol/L AlCl3处理下,设计0、4、8、12、16、24、36、48和72 h共9个时间梯度,对GmGST7的表达模式进行分析。过表达GmGST7基因遗传转化拟南芥,鉴定阳性植株,并对转基因株系进行耐酸铝表型验证、氧化水平测定、耐酸铝标志基因及下游基因的表达分析。【结果】GmGST7基因位于大豆第7号染色体,序列全长为1128 bp。该基因含有2个外显子和1个内含子,2个外显子分别编码GST高度保守的N端和不保守的C端;GmGST7基因编码226个氨基酸,编码的蛋白为大豆GST蛋白的tau类家族成员,定位于细胞质和细胞核中;GmGST7基因在大豆根、茎、叶、花和幼荚中均有表达,且在根中的表达量最高;GmGST7基因在50μmol/L AlCl3处理24 h时表达最高;AlCl3处理后,野生型拟南芥相对根伸长显著低于转基因株系的,野生型拟南芥氧化水平高于转基因株系的,耐酸铝标志基因和下游基因的表达量在转基因株系中较高。【结论】GmGST7基因属于大豆GST tau类家族成员,在细胞核和细胞质中行使功能,呈组成型表达模式,且在大豆根中表达最高,对酸铝胁迫响应显著;GmGST7过表达通过激活酸铝胁迫标志基因及其下游基因的表达提高拟南芥的酸铝耐受能力。

18份大豆品系在中国南方地区的适应性评价

本试验采用国家大豆改良中心广东分中心的18个大豆品系,在广东广州、广西南宁和湖南长沙三个不同的试验地点种植,分析不同生态环境对大豆农艺性状、产量、品质的影响,并对供试品系在不同区域的适应性进行评价,结果表明:18份品系在长沙种植的平均株高、平均百粒重、平均单株粒重均显著高于广州与南宁,平均粗蛋白含量显著高于广州,但平均粗脂肪含量显著低于其他两地;通过相关性分析发现,品系粗蛋白含量与粗脂肪含量在个试验地点均呈显著负相关,说明二者难以兼顾,育种过程种应有取舍或平衡;三个试验点影响产量的主要性状均包含单株粒数,其他性状如株高、百粒重等对产量的影响在三地并不相同,培育南方广适高产的大豆品种,应首先关注单株粒数;对产量和品质综合评价,粤夏110和粤夏114在广州、南宁、长沙三地产量高、品质好,表现稳定,适应性好。

光质对菜用大豆苗期光形态建成及根冠比的影响

发光二极管(LED)已被广泛地应用于温室栽培作物及科研的光源。本研究以上海青、华夏7号和浙鲜豆5号3个品种菜用大豆品种为供试材料,采用LED植物生长灯为光源,研究相同光照强度(180±5μmol·m^(-2)·s-1)下,不同光质对菜用大豆光形态建成和根冠比的影响。结果表明:与白光相比,以红光为主的光质处理显著提高了菜用大豆的株高和下胚轴长度,随着组合光中红光比例的增加,植株的复叶面积逐渐增大;蓝光及以蓝光为主的组合光明显促进了根系的生长;相对于单色光组合光能提高菜用大豆地上部生物量、地下部生物量及根冠比;菜用大豆对光质的响应存在着品种间差异。

大豆品种特性对腐竹产量及品质的影响

【目的】大豆(Glycine max L.)含有丰富的植物蛋白和油脂,因而成为一种具有较高经济价值的作物。探究不同大豆品种的腐竹加工产量及不同大豆品种生产的腐竹在蛋白质、油分、可溶性糖、异黄酮之间的相关性,为制作生产高异黄酮腐竹提供参考依据。【方法】采用来自黑龙江和广东大豆产区的品种24份,用同一工艺制作腐竹,然后用凯氏定氮法测定大豆和腐竹中蛋白质,用索氏抽提法测定油分,用蒽酮比色法测定可溶性糖,用高效液相色谱法测定大豆和腐竹中的异黄酮含量。【结果】不同品种在腐竹产量和品质方面都存在明显的差异,大豆品种华夏8号制得腐竹产率最高,达到60.50%;其次为品种华春2号,为52.44%,这两个品种是制作腐竹的理想品种;此外,绥农37、华春6号和黑河43的腐竹生产率分别达到了48.59%、48.37%和47.91%,也是产率比较高的品种。相关分析结果表明,腐竹产量与大豆中蛋白质含量呈显著正相关(r=0.598**),与大豆中的可溶性糖呈负相关(r=-0.423*)。腐竹蛋白质含量、油分含量及异黄酮含量3个性状都分别与大豆种子对应的性状呈极显著正相关(r分别为0.700**、0.537**和0.879**);腐竹可溶性糖含量与大豆可溶性糖含量呈显著正相关(r=0.441*)。腐竹中的蛋白质含量与大豆可溶性糖含量呈极显著负相关(r=-0.519**)。腐竹中的油分含量与大豆中蛋白质呈极显著负相关(r=-0.889**),与大豆中可溶性糖和异黄酮含量呈极显著正相关(r分别为0.614**和0.574**);腐竹中异黄酮含量与大豆中蛋白质含量呈极显著负相关(r=-0.589**),与大豆中可溶性糖含量呈极显著正相关(r=0.568**)。【结论】大豆品种的腐竹产率和主要品质性状存在显著差异,其中,华夏8号、华春2号是制作腐竹的高产品种,大豆品种的品质特性决定了腐竹的品质特性,其主要由大豆品种的遗传特性决定的。

废弃矿区大豆根际土壤细菌群落对施肥方式的响应

【目的】从细菌多样性和组成结构角度评估化学肥料与硅酸钠或蚯蚓结合施用对废弃矿区大豆根际细菌群落的影响,为日后矿区复垦和生态恢复提供理论依据。【方法】在广东省梅州市黄畲村废弃矿区种植大豆‘华春9号’,试验设置4个施肥处理:氮磷钾肥结合硅酸钠(3NPK+S和5NPK+S),氮磷钾肥结合蚯蚓20条·m−2(3NPK+E和5NPK+E),以不施肥为对照(3CK和5CK)。在施肥后3和5个月时对大豆根际土壤进行采样。通过提取土壤细菌总DNA、16S rRNA测序和测定土壤基本化学性质,探究该矿区大豆根际细菌群落多样性和结构对不同施肥处理的动态响应过程。【结果】与3CK相比,3NPK+E处理显著增加了Chao1和Shannon指数;与5NPK+E处理相比,5NPK+S处理显著增加了Shannon指数。主坐标分析(PCoA)表明,不同处理下的根际细菌群落组成在操作分类单元(OTU)水平上出现显著分离。从属的相对丰度来看,Uncultured_f_Anaerolineaceae是3NPK+S、3NPK+E、5NPK+S、5NPK+E处理土壤中的优势菌属。基于Mantel检验和典型关联分析(CCA)显示,土壤有机质、有效磷和全碳含量对5NPK+E处理的根际细菌群落有显著影响。【结论】施加NPK+S和NPK+E肥料均能有效提高矿区土壤细菌多样性,从而间接对土壤修复起促进作用。施肥方式和处理时间均会显著影响细菌群落组成,并且施肥处理时间越长对细菌群落组成的影响越明显。Uncultured_f_Anaerolineaceae富集在施肥处理下的土壤中,这类细菌可能在促使碳转化的过程中起重要作用,同时还具有修复土壤污染的潜力。NPK+E施用增加了有机质含量,对细菌群落的生长有促进作用。

大豆遗传转化方法及再生体系研究进展

转基因技术作为现代生物技术之一,在基因功能研究和转基因育种方面取得重要进展。大豆基因组测序之后,大豆功能基因组学发展迅速,挖掘控制重要性状的基因用于转基因育种受到广泛关注。随着转基因大豆新品种培育重大专项的实施,我国建立了基因克隆、遗传转化、功能研究、转基因品系安全评价等转基因育种研究技术体系和监管体系。其中,转基因大豆遗传转化技术体系的研究取得了较快的进展,主要集中在高效、稳定遗传转化再生体系的建立和优化,大豆遗传转化方法的探索和优化等方面。对大豆遗传转化体系、转化方法和转化效率等因素进行阐述,可为大豆基因功能研究和转基因新品种培育相关研究提供参考。

Fine mapping and genetic analysis of resistance genes,Rsc18,against soybean mosaic virus

Soybean mosaic virus(SMV) affects seed quality and production of soybean(Glycine max(L.) Merr.) worldwide.SC18 is one of the dominant SMV strains in South China,and accession Zhonghuang 24 displayed resistance to SC18.The F_(1),F_(2) and 168 F_(11) recombinant inbred lines(RILs) population derived from a hybridization between Zhonghuang 24(resistant,R) and Huaxia 3(susceptible,S) were used in this study.According to the segregation ratios of the F_(2) generation(3 R:1 S) and the recombinant inbred lines(RILs) population(1 R:1 S),one dominant locus may regulate the resistance to SC18 in Zhonghuang 24.By using composite interval mapping(CIM),Rsc18 was mapped to a 415.357-kb region on chromosome 13.Three candidate genes,including one NBS-LRR type gene and two serine/threonine protein type genes,were identified according to the genetic annotations,which may be related to the resistance to SC18.The q RT-PCR demonstrated that these genes were up-regulated in the R genotype compared to the control.In conclusion,the findings of this research enhanced the understanding about the R genes at the Rsc18 locus.Moreover,our results will provide insights for designing molecular markers to improve marker-assisted selection and developing new varieties with resistance to SC18.

广东大豆产业现状及原料需求分析

广东是我国大豆加工大省,大豆食品加工企业经济效益突出,仅酱油一个产品纯利润就高达100亿元以上。2006年以后广东大豆育种取得了突破,先后审定了25个新品种,大豆平均公顷产量达到2.6吨。广东大豆食品加工年大豆原料需求约200万吨,其中酱油和豆酱原料约120万吨。但是,广东生产大豆少,主要原料依靠在国内各省进口。所以,广东省大豆产业的发展与黑龙江、内蒙古、吉林、长江流域、黄淮海各省大量的原料进口和建立广东大豆产业技术创新联盟有密切的关系。

Identification and validation of stable and novel quantitative trait loci for pod shattering in soybean[Glycine max(L.)Merr.]

Pod shattering is an important domesticated trait which can cause great economic loss of crop yield in cultivated soybean.In this study,we utilized two recombinant inbred line populations(RILs,CY,Huachun 2×Wayao;GB,Guizao 1×B13)to identify quantitative trait loci(QTLs)associated with pod shattering in soybean across multiple environments.A total of 14 QTLs for pod shattering were identified in the two RIL populations,which had LOD scores ranging from 2.64 to 44.33 with phenotypic variance explanation(PVE)ranging from 1.33 to 50.85%.One QTL qPS16-1,located on chromosome 16,included a well-known functional gene Pod dehiscence 1(Pdh1)that was reported previously.Ten new putative QTLs were validated in two RIL populations,and their LOD scores were between 2.55 and 4.24,explaining 1.33 to 2.60%of the phenotypic variation.Of which four novel QTLs(qPS01-1,qPS03-2,qPS05-1,and qPS07-1)could be detected in two environments where nine genes had specific changes in gene expression.Although the nine genes may have significant effects on pod shattering of soybean,their detailed functions still need to be further explored in the future.The results of this study will facilitate a better understanding of the genetic basis of the pod shattering-resistant trait and benefit soybean molecular breeding for improving pod shattering resistance.

Effect of Inoculation Rhizobium and Response of Soybean-Rhizobium System to Insoluble Phosphate

In order to produce efficiently symbiotic system in acid soils which could be used for the recovery of insoluble phosphate soil and improvement of the fertility, etc., host range of Rhizobium was isolated from the root nodules of wild and cultivated soybean varieties growing in south China, evaluated the effects of Rhizobium on enhance nitrogen fixing activities, phosphorus(P), nitrogen(N) uptakes, growth and yield of soybean under pot and field conditions. The results showed that combined Rhizobium inoculation was more effective than individual Rhizobium inoculation. The combined Rhizobium of CW54 strains had the highest effect, nodule number, nodule weight and nodule nitrogenase activity were markedly stimulated as compared to those of the control U110, respectively. The nitrogen contents in the plants treated with CW54 strains were significantly increased by 43.2% as compared to those of the control U110 in Al-P soil, but statistical significance was not observed in Fe-P soil. By contrast, inoculation with CW54 significantly enhanced phosphorus uptake of plant in Fe-P soil than that in Al-P soil. The effects of inoculation of CW54 strains in a field condition were similar to those of in a pot condition. The total biomass and yield in the plants treated with CW54 strains were significantly increased by 86.4% and 51.4% as compared to those of the control at the time of soybean podding in a field condition, respectively. These results suggested that inoculation Rhizobium was beneficial for soybean nutrient uptake, growth and yield under insoluble phosphate soil conditions.

Effects of Ecological Conditions of the Northeast of China on Protein，Oil and Protein＋ Oil Content in Soybean Seeds

Twenty five soybean strains of similar growth period were grown at eight locations simultanously in three northeast provinces of China.Most of the experimental sites differed markedly in temperature,precipitation, soil fertility and length of growing season.Protein and oil contents were detelmined to study the effects of ecological conditions on soybean chemical composition. The results showed that the average protein and protein + oil contents were higher at the south locations and lower at north locations, but there was no consistent variation for oil content,and there was no significant difference among most of the sites. Oil content of strains was highest when planted in Jiling. Combined analyses of variance indicated that there existed significant genotype,location and G×E interactions effects. F values of G×E interactions for all traits,although significant at 0.01 level of probability, were extremely smaller than those for location and genotype.This suggested that G×E interaction effects were relatively less important than both genotyp and location effects.Phenotypic correlations of the performance of genotypes,on single ehtry -basis,among eight locations were all positive for protein and protein+ oil and most of correlation coefficients were significant at 0.01 level of probability.This indicated that the relative performance of genotypes at each location was similar. In contrast,most of the correlation coefficients for oil content among the eight locations were not significant.Therefore,G×E interactions should be considered in breeding for high oil soybeans,as it may lead to the instability of genotypes and affect the rankings of genotypes when soybeans arc grown in diverse environments.