8株优良大豆根瘤菌与不同地区27个大豆主栽品种的匹配性研究

大豆是我国重要的油料作物和高蛋白粮饲兼用作物,具有与根瘤菌共生固氮的能力。近十几年来,我国大豆品种更新快,导致大豆根瘤菌株与新品种匹配能力差、接种效果不明显。筛选与主栽大豆品种匹配性好、固氮效率高的广谱性优良菌株,可为针对性的施用大豆根瘤菌接种剂提供菌种资源和方案。选取本实验室前期分离保存的6个优良快生型大豆根瘤菌株和2个慢生型大豆根瘤菌株,在砂培盆栽条件下与不同地区的27个大豆主栽品种进行匹配试验。测定分析了植株地上部分生物量、高度、根瘤数量、根瘤生物量和根瘤固氮酶活指标。结果表明:不同大豆根瘤菌之间结瘤固氮能力存在极显著的差异;供试根瘤菌均能够与国内24个大豆品种结瘤,广谱性较好;植株地上部分高度、根瘤数量、根瘤生物量和根瘤固氮酶活与地上部分生物量呈显著相关;大部分接种根瘤菌后的植物生物量显著高于CK;HN01、GR3、HH29、HH103匹配性和固氮效率均不逊色于USDA110,具有在东北地区、黄淮海地区、长江流域、东南地区推广的潜能;从品种来看,中豆39、BD2、天隆1号与8株供试大豆根瘤菌的匹配接种表现出高生物量特点。此外,本文还筛选出大豆-大豆根瘤菌的表型最佳匹配组合中豆39-GR3,适合长江流域地区;同样筛选到东北地区、黄淮海地区、东南地区最佳匹配组合,分别是HN01-辽豆14、HN01-徐豆14、HN01-BD2。本文初步建立了优良根瘤菌与大豆主栽品种的匹配关系,为在田间试验中进一步筛选和应用这些优良菌株提供了材料和指导。     

豆科作物氮素高效利用机制研究进展

工业氮肥的使用一定程度上满足了农作物高产的需求,但过量施用带来严重的环境生态问题,如何利用生物固氮提供的绿色高效氮素部分代替工业合成氮肥是当前研究的热点。虽然共生固氮体系效率较高,但肥料氮和根际微生物群落可能对结瘤固氮体系的利用产生消极影响。为此,本文从制约共生固氮体系高效利用的关键因素,即土壤有效氮含量及根际微生物群落入手,梳理了豆科植物-根瘤菌共生体系的建立及其对外源氮的响应以及根际微生物群落与根瘤菌共生效率的关系,总结分析了氮素高效利用的生理分子机制,通过优化调控技术提高氮素利用率,为豆科作物氮素高效利用提供理论支持。     

减氮、加菌、改善土壤物理性状提高大豆固氮能力

氮是大豆生长必须的营养元素之一。大豆是豆科作物,具有共生固氮能力,据估算东北黑土区大豆共生固氮潜力在150～200kg/hm2之间,同时大豆的共生固氮能力受多种因素的影响。土壤中的根瘤菌对氮的响应表现为使用过多的氮肥会抑制根瘤菌的繁殖生长,但少量地使用化肥有利于提高大豆的共生固氮能力。改善土壤物理性状能够增加土壤中的根瘤数量,进而提高大豆的共生固氮能力。基于上述事实我们提出了"减氮、加菌、改善土壤物理性状"的模式,该模式将大豆产量提高了15.3%,氮肥节约了33.3%。     

大豆根瘤菌HH103ΩNopAAΩNopD双突变体构建及结瘤能力鉴定

大豆—根瘤菌的共生模式为大豆提供了丰富的氮源。根瘤菌Ⅲ型效应因子是影响共生结瘤的关键信号分子之一,通过对HH103ΩNopAA与HH103ΩNopD的结瘤鉴定表明两个突变体对绥农14和ZYD00006的宿主亲和性不同,而后通过三亲杂交方法在HH103ΩNopAA基础上构建HH103ΩNopAAΩNopD的双突变体。利用绥农14与野生豆ZYD00006对突变体进行结瘤鉴定,HH103ΩNopAAΩNopD的根瘤数与根瘤重与HH103ΩNopAA无显著性差异。而后利用前期收集的100份大豆资源进行结瘤鉴定,分析NopAA与NopD的突变在不同遗传背景下的大豆品种的结瘤特性,绝大多数品种根瘤数明显减少,部分品种根瘤数在接种单突变体与双突变体存在显著差异。该研究为后续解析NopAA与NopD的共生信号传导以及根瘤菌与大豆品种亲和性提供新的思路。同时可以根据基因型的差异,筛选与不同根瘤菌均有较高亲和性的大豆品种,为进一步培育高结瘤、高固氮的大豆品种提供支撑。     

大豆硫营养

水培矮脚早大豆研究表明；大豆生育适宜的硫量为０．１ｍＭｏｌ．Ｓ．Ｌ＾－１，能促进干物质积累。不同硫水平营养器干物质分配率（ＰＲＤＭ）均是成熟期低于前期。种子中ＰＲＤＭ与硫量负相关。器官中硫含量高低依种子，叶，茎，根，荚皮顺序递减。硫有促进共生固氮功能，荚期茎部酰脲及酰脲丰度以０．５ｍＭｏｌ．Ｓ．Ｌ＾－１处理最高。缺（低）硫处理器官酰脲与硝态氮增加。花期功能一非功能叶全硫量分别低于０．１２％，０．０     

大豆共生固氮能力对土壤无机氮浓度的响应与调控

通过对国内外大豆共生固氮总量及占大豆一生需氮量的比例,以及大豆共生固氮能力对土壤无机氮浓度的响应的综述,提出了如何提高大豆共生固氮能力的途径。明确了协调土壤环境氮浓度和大豆根瘤固氮之间的关系,是充分发挥大豆共生固氮潜力和提高产量的关键所在。     

大豆-根瘤菌的固氮作用

报道了我国主要大豆产区根瘤菌和寄主品种固氮资源的分布、主要类型、固氮作用。我国多年连续种植的大豆的田块 ,根瘤菌菌数每克干土一般超过 10 4 个 ,大部分零星隔年种植和不常种植大豆的田块根瘤菌数每克干土低于 10 4 个。多数根瘤菌土著者能与大豆栽培品种有效结瘤 ,但主要是中、低效固氮者。在自然条件下 ,供试的春大豆中高效固氮者占 31.7% ,供试的夏大豆高效者占 6 4.3% ;人工接种根瘤菌 ( 113 - 2 ) ,春豆中高效固氮者占 40 .9% ,夏豆占 5 3.6 %。我国的栽培大豆是较好的耐高氮共生固氮资源 ,评价出 2个在 5mMNO-3 条件下结瘤的品种。     

高产高蛋白大豆与高效固氮根瘤菌和菌根菌最佳联合共生体筛选

大豆——根瘤菌——菌根真菌联合共生对大豆生产具有重要意义,接种根瘤菌能扩大氮肥来源、接种菌根真菌有助于作物从土壤中吸取更多的磷素,应用这两项生物技术可改善大豆氮磷营养、提高土壤肥料利用率,是作物走向自身从空中取氮,向土壤索磷的新方向。 土壤中存在着大量的土著根瘤菌和菌根真菌,要选择高效根瘤菌和菌根菌与之共生,     

Nitrogen Fixation Establishment during Initial Growth of Grain Legume Species

Atmospheric nitrogen fixation as a result of the symbiosis between bacteria and legume species, can result in major advantages in providing host plants with organic nitrogen. The objective of the present study was to evaluate the physiological potential during early seedling development for initiation of nodulation and nitrogen fixation activity of four grain legumes species: soybean [Glycine max (L.)], cowpea [Vigna unguiculate (L.) Walp], common bean [Phaseolus vulgaris (L.)], and peanut [Arachis hypogaea (L.)]. Seedlings were grown on a hydroponic solution so that nodule development could be readily observed until about 3 weeks after germination. Nodules developed in all cases. Acetylene reduction activity (ARA) by soybean and cowpea was also found early in seedling development. In contrast, peanut and common bean showed little or no development of ARA during seedling development. The results provided insight into differences in physiological potential among grain legumes in establishing symbiotic nitrogen fixation during crop establishment. These results indicate those species/cultivars that are candidates for readily establishing nitrogen fixation activity during the seedling stage of plant development.     

不同施氮量及施氮方式对大豆根瘤生长及产量的影响

以宁夏地区主推品种中黄30为主要研究对象,采用再裂区试验设计,通过接种高效根瘤菌,探讨根瘤菌、施氮量和施氮方式对大豆根瘤干重、根瘤数量以及大豆产量的影响。结果表明:施氮量显著影响大豆结瘤和大豆产量;各处理随着每次施氮量的增加,根瘤干重和根瘤数量的变化趋势均为先增加后下降;产量表现为分次施氮各处理产量一次性施氮各处理产量不施氮(B1)产量;接种根瘤菌处理较不接种根瘤菌处理的根瘤数量及大豆产量显著增加;是否接种根瘤菌与施氮量互作、施氮量与施氮方式互作以及是否接种根瘤菌、施氮量和施氮方式互作对大豆产量均产生极显著影响,中黄30产量最佳组合是A2B3C2(接种根瘤菌、施氮量在75 kg·hm~(-2)且分次施氮)的前提下,产量最高,达到6 020.37 kg·hm~(-2),较不施氮增产8.36%。     

铵硝配比对大豆生长及结瘤固氮的影响

通过水培方法在5种不同铵硝配比下,研究不同氮形态及接种根瘤菌对大豆生长、结瘤固氮的影响及其生理机制。结果表明,不同铵硝配比对大豆植株生长、产量、生物固氮及氮效率等影响显著,随硝氮比例增加,植株生物量、籽粒产量、结瘤固氮及氮效率等指标均呈现先增加后降低的趋势,以铵硝比25:75处理效果最佳,总体表现为铵硝比25:75≥0:100>50:50>75:25>100:0。较高铵硝比（≥50:50）严重抑制了大豆根系的伸长生长、根瘤的形成及其固氮酶的活性,从而降低了生物固氮的贡献率,最终导致大豆产量及氮素效率的降低。接种根瘤菌显著增加大豆产量和籽粒含氮量,并与铵硝比之间存在协同效应。铵硝比为25:75时接种根瘤菌,氮效率最高,最有利于大豆生长及籽粒产量的形成。研究结果对优化大豆水培营养液配方以及指导农田生产中氮素合理施用均有较强的借鉴作用。     

大豆分子育种研究进展

分子育种技术的研究和应用是近十余年来世界大豆生产发展的重要科技动力。2011年,国内外在大豆分子育种相关领域取得了新的进展。在分子标记开发与辅助育种方面,发掘出与产量、发育、品质、抗病和抗逆等性状相关的新的分子标记或QTL;在新基因挖掘方面,克隆了与光周期反应、共生固氮、品质及抗逆性相关的基因,并分析了其功能。在大豆转基因育种方面,遗传转化体系的优化取得了新的进展,转化效率有所提高,同时对一些功能基因(包括来自其它物种的一些基因)进行了功能评价和育种利用价值评估。转基因大豆继续保持良好的发展势头,并且出现一些新的发展方向。该文对这些研究进展进行了综述,并对今后的发展趋势进行了展望。     

根瘤细菌的生物学多样性和细菌—宿主改良

针对我国主要豆科作物大豆和花生的研究成果，并结合本身研究工作内容，（1）总结了与宿主共生的根瘤菌生物学多样性，包括大、小样本的土著根瘤菌群体数量、分离频率、类型与分布及其菌株-宿主共生混交性与亲和性；（2）评述了根瘤菌-宿主植物共生体双边固氮改良，包括优良菌株的选育、宿主品种资源共生特性的评价与利用及特异性状的选择；（3）讨论了我国这一类共生体资源的丰富性以及有必要进一步加大力度研究有益资源的开发和利用的问题。     

Rhizobial Inoculants for Legume Crops

Summary

Legumes are an important source of protein for humans and livestock. Legumes have also been used for soil improvement for centuries because of their N and non-N rotational benefits to non-legume crops. The N benefits include N₂ fixation and mineralization, sparing of soil inorganic N, and reduced immobilization of soil inorganic N. The non-N benefits include breaking pest cycles, improvement of soil structure, and the nutritional and disease-control effects of endophytic rhizobia. Therefore, optimizing the legume-Rhizobium symbiosis is important, and it can be done by selecting or modifying either (or both) symbiotic partner(s) for desirable traits related to N₂ fixation. Rhizobium strains can be selected or genetically modified for traits like N₂ fixation potential, nodulation competitiveness, persistence in soil, compatibility with inoculant carriers, and tolerance to environmental stress factors. Legume genotypes can also be selected, bred or genetically modified for N₂ fixation potential, restricted or preferential nodulation, and tolerance to nitrate and environmental stress factors. When choosing prospective strains or legume genotypes for a particular environment, time and resources can be saved by realizing that the most adaptable rhizobia or legume genotypes are usually those isolated from similar environments. Inoculant delivery methods also affect N₂ fixation. Soil inoculation, particularly with granular inoculants, seems to be often better and never worse than seed inoculation for initiating nodulation and N₂ fixation. Use of pre-inoculated seeds eliminates the seed inoculation operation, but Rhizobium numbers in pre-inoculated seeds tend to be lower than those in traditional inoculant products. Therefore, the time saved by using pre-inoculated seeds should be weighed against the possibility that crop yields may be lower if insufficient Rhizobium numbers are delivered. Until tools for genetic modification of rhizobia or legumes to suit specific requirements are commonly used, N₂ fixation can be enhanced by adopting practices like choosing the best combinations of Rhizobium strains and legume genotypes, the best inoculant formulation and delivery methods, optimum inoculation rates, and providing favourable growing conditions for the crop.     

高效快生型大豆根瘤菌的筛选及分子标记

采用黑龙江省不同土壤类型对20株不同地区来源的根瘤菌和5个大豆品种(品系)进行接种效果的研究,旨在使筛选结果尽量接近实际生产环境,筛选出符合实际效果的高效共生快生型菌株.结果表明,大豆与根瘤菌之间表现出共生效果的多样性.针对不同土壤相应的大豆品种有其高效匹配的快生根瘤菌.初步确定了以AFLP技术作为高效菌株的分子标记,为进一步研究高效菌株在土壤中定植以及与土壤微生物、植物之间的生态学研究提供依据.     

大豆种质资源共生固氮特性评价及遗传初步研究

以我国２２个省５９０份主要栽培大豆品种为材料,评价春、夏、秋大豆遗传资源结瘤固氮性状,并配制杂交组合进行初步遗传分析,结果显示,１）春夏秋大豆类型间、品种间结瘤固氮性状差异极显著。同类型不同产区的大豆品种共生固氮性状差异显著,以长江流域及其以南产区,尤以湖北、江苏省品种最佳。２）与共生固氮性状密切相关的１０项参数变异范围以夏大豆最大,并鉴定出单株结瘤数达２３８．８个,根瘤重达０．５９ｇ、固氮量达１     

Drought tolerance and yield increase of soybean resulting from improved symbiotic N2 fixation

Drought is by far the most important environmental factor contributing to crop yield loss, especially in soybean [Glycine max (L.) Merr.] where symbiotic fixation of atmospheric nitrogen (N₂) is sensitive to even modest soil water deficits. Decline of N₂ fixation with soil drying causes yield reductions due to inadequate N for protein production, which is the critical seed product. In this paper, we present a combined physiological and breeding research effort to develop soybean lines that have diminished sensitivity of N₂ fixation to drought. A preliminary physiological screen was used to identify lines that potentially expressed N₂ fixation drought tolerance. One hundred progeny lines derived from a cross between Jackson, a cultivar proven to have N₂ fixation tolerance to drought, and KS4895, a high-yielding line, were tested in the screen. Seventeen lines were identified for subsequent yield trials in moderate- and low-yielding rainfed environments. Two lines, found to have higher yields than commercial checks in these environments were then tested in the greenhouse for their N₂ fixation activity in drying soil. Nitrogen fixation activity was found to persist at lower soil water contents than exhibited by the sensitive parent. These two soybean lines offer a genetic resource for increased yields under rainfed conditions as a result of decreased sensitivity of N₂ fixation to water deficit.     

Model legumes contribute to faba bean breeding

Faba bean is an excellent candidate crop to provide nitrogen input into temperate agricultural systems. However, its growth is hampered by several factors including environmental stresses and the presence of anti-nutritional factors. To solve these limitations, breeding programs have been initiated that were successful for monogenic traits but not so for multigenic traits. The large genome size of faba bean has slowed down breeding processes. Several other legumes have emerged as model legumes including Medicago truncatula, Lotus japonicus, Glycine max and Pisum sativum. The establishment of these models has already boosted our understanding of important processes such as the nitrogen-fixing symbiotic interaction. The high level of synteny and collinearity existing between legumes makes possible the transfer of key knowledge from model legumes to faba bean. Here we review the most recent knowledge gained from model legumes on grain quality, resistance to biotic and abiotic stresses, nitrogen-fixing symbiosis and how this knowledge can be employed for faba bean breeding.     

大豆结瘤固氮的分子生理研究

有关豆科作物共生固氮的研究已有170 a的历史。而豆科结瘤固氮的分子生理研究,仅仅是最近30 a的事情。从影响大豆结瘤固氮的生理因子角度,评述了豆科作物的结瘤自动调控机制,进一步在分子水平上阐述了生长素、硝态氮、黄酮类等生理调控物质对大豆结瘤固氮的影响。生长素调节根瘤菌侵染位点和根瘤形成后的生长;可通过控制结瘤的角度来减少土壤硝态氮对大豆结瘤固氮能力的抑制;黄酮类物质在根瘤发育和结瘤基因诱导中起重要作用;乙烯抑制结瘤信号的前期过程,调整根瘤形成的空间分布。详细、系统地从作物-微生物之间关系出发,研究引起豆科作物根瘤形成和固氮系统的整体信号传导、蛋白和代谢过程,是未来大豆共生固氮分子生理研究应该注重的方向。     

Molybdenum-enriched soybean seeds enhance N accumulation, seed yield, and seed protein content in Brazil

High soybean yields require large amounts of nitrogen (N), which can be obtained mainly from symbiotic N₂ fixation. However, the efficiency of this biological process can be limited by micronutrient deficiencies, especially of molybdenum (Mo). In Brazil, soybean generally responds positively to fertilization with Mo in soils of low fertility and in fertile soils depleted of Mo due to long-term cropping. The micronutrient can be supplied by seed treatment, however toxicity of Mo sources to Bradyrhizobium strains applied to seed as inoculant has been observed, resulting in bacterial death and reductions in nodulation, N₂ fixation and grain yield. Therefore, use of seeds enriched in Mo could be a viable alternative to exterior seed treatment, allowing elite inoculant strains of Bradyrhizobium to sustain high rates of biological N₂ fixation. We demonstrated the feasibility of producing Mo-rich seeds of several soybean cultivars, by means of two foliar sprays of 400 g Mo ha⁻¹ each, between the R3 and R5 stages, with a minimum interval of 10 days between sprays. As a result of this method, considerable increases in seed-Mo content were obtained, of as much as 3000%, in comparison to seeds obtained from plants which received no Mo. In field experiments performed in soils with low N content and without any N-fertilizer supply, inoculation of Mo-rich seeds produced plants with increased N and Mo contents in the grain and higher yields of total N and of grain. In most cases, Mo-rich soybean seeds did not require any further application of Mo-fertilizer.