Nodulation Capacity of Argentinean Soybean (<i>Glycine max</i>L. Merr) Cultivars Inoculated with Commercial Strains of <i>Bradyrhizobium japonicum</i>

The purpose of this research was to evaluate the nodulation potential of 31 Argentinean soybean commercial cultivars. Those with the highest nodulation capacity response developed twice the amount of nodules than the low nodulating ones, which is the variation contained in soybean genotypes. Furthermore, this was not due to bacterial promiscuity, since the response was independent of the bradyrhizobia strain inoculated. The ability of cultivars to develop a larger number and biomass of nodules was unrelated with the maturity group they belong to and also was not a response to quorum sensing effects. Our results suggest that breeding programs can be aimed at improving the nodulation capacity of soybean and that cultivars from different maturity groups can be a source of nodulation QTLs.

A high-resolution transcriptomic atlas depicting nitrogen fixation and nodule development in soybean

Although root nodules are essential for biological nitrogen fixation in legumes,the cell types and molecular regulatory mechanisms contributing to nodule development and nitrogen fixation in determinate nodule legumes,such as soybean(Glycine max),remain incompletely understood.Here,we generated a single-nucleus resolution transcriptomic atlas of soybean roots and nodules at 14 days post inoculation(dpi)and annotated 17 major cell types,including six that are specific to nodules.We identified the specific cell types responsible for each step in the ureides synthesis pathway,which enables spatial compartmentalization of biochemical reactions during soybean nitrogen fixation.By utilizing RNA velocity analysis,we reconstructed the differentiation dynamics of soybean nodules,which differs from those of indeterminate nodules in Medicago truncatula.Moreover,we identified several putative regulators of soybean nodulation and two of these genes,GmbHLH93 and GmSCL1,were as-yet uncharacterized in soybean.Overexpression of each gene in soybean hairy root systems validated their respective roles in nodulation.Notably,enrichment for cytokinin-related genes in soybean nodules led to identification of the cytokinin receptor,GmCRE1,as a prominent component of the nodulation pathway.GmCRE1 knockout in soybean resulted in a striking nodule phenotype with decreased nitrogen fixation zone and depletion of leghemoglobins,accompanied by downregulation of nodule-specific gene expression,as well as almost complete abrogation of biological nitrogen fixation.In summary,this study provides a comprehensive perspective of the cellular landscape during soybean nodulation,shedding light on the underlying metabolic and developmental mechanisms of soybean nodule formation.

Coordinated responses of leaf and nodule traits contribute to the accumulation of N in relay intercropped soybean

Maize(Zea mays L.)-soybean(Glycine max L.Merr.)relay intercropping provides a way to enhance land productivity.However,the late-planted soybean suffers from shading by the maize.After maize harvest,how the recovery growth influences the leaf and nodule traits remains unclear.A three-year field experiment was conducted to evaluate the effects of genotypes,i.e.,supernodulating(nts1007),Nandou 12(ND12),and Guixia 3(GX3),and crop configurations,i.e.,the interspecific row spacing of 45(I45),60(I60),75 cm(I75),and sole soybean(SS),on soybean recovery growth and N fixation.The results showed that intercropping reduced the soybean total leaf area(LA)by reducing both the leaf number(LN)and unit leaflet area(LUA),and it reduced the nodule dry weight(NW)by reducing both the nodule number(NN)and nodule diameter(ND)compared with the SS.The correlation and principal component analysis(PCA)indicated a co-variability of the leaf and nodule traits in response to the genotype and crop configuration interactions.During the recovery growth stages,the compensatory growth promoted soybean growth to reduce the gaps of leaf and nodule traits between intercropping and SS.The relative growth rates of ureide(RGR_U)and nitrogen(RGR_N)accumulation were higher in intercropping than in SS.Intercropping achieved more significant sucrose and starch contents compared with SS.ND12 and GX3 showed more robust compensatory growth than nts1007 in intercropping.Although the recovery growth of relay intercropping soybean improved biomass and nitrogen accumulation,ND12 gained a more significant partial land equivalent ratio(pLER)than GX3.The I60 treatment achieved more robust compensation effects on biomass and N accumulation than the other configurations.Meanwhile,I60 showed a higher nodule sucrose content and greater shoot ureide and N accumulation than SS.Finally,intercropping ND12 with maize using an interspecific row spacing of 60 cm was optimal for both yield advantage and N accumulation.

Identification of Rhizobia Isolated from Nodules of Mexican Commercial Soybean Varieties

Rhizobia, crucial for nitrogen fixation in leguminous plants, play a vital role in soybean cultivation. This study, conducted in Mexico, a major soybean importer, aimed to identify bacteria from nodules of five soybean varieties in high-production regions. Multilocus sequence analysis (MLSA) was employed for enhanced species resolution. The study identified six Bradyrhizobium species: Bradyrhizobium japonicum USDA 110, Bradyrhizobium japonicum USDA 6, Bradyrhizobium elkanii USDA 76, Bradyrhizobium neotropicale, Bradyrhizobium lablabi, and Bradyrhizobium icense. Bradyrhizobium japonicum USDA 110 predominated in the soils, displaying symbiotic preference for the Huasteca 400 variety. However, phylogenetic analysis didn't reveal a clear association between strains, soil, and soybean variety. This research sheds light on the diversity of rhizobia in Mexican soybean cultivation, contributing to the understanding of symbiotic relationships in soybean production systems.

Nodule Formation and Development in Soybeans(Glycine max L.) in Response to Phosphorus Supply in Solution Culture

Phosphorus (P) is necessary for growth and nitrogen fixation, and thus its deficiency is a major factor limiting legume production in most agricultural soils. The effect of phosphorus supply on nodule development and its role in soybeans (Glycine max L.) was studied in a nutrient solution. Plants were inoculated with Bradyrhizobium japonicum and grown for 35 days in a glasshouse at a day and night temperature of 25℃and 15℃, respectively. Although increasing P supply increased the concentrations of P and N in the shoots and roots, the external P supply did not significantly affect the P concentration in the nodules, and the N fixed per unit nodule biomass decreased with increasing P supply. The nitrogen content in the shoots correlated well with the P content (r = 0.92**). At an inoculation level of 102 cells mL-1, the P supply did not affect the number of nodules; however, at inoculation levels of 103.5 and 105 cells mL-1, increasing P supply increased both the number and size of nodules. Irrespective of the inoculation level, increasing P supply increased the nodule biomass relative to the biomass of the host plant. It is suggested that the P deficiency specifically inhibited the nodule development and thereby the total N2 fixation.

Mechanisms underlying key agronomic traits and implications for molecular breeding in soybean

Soybean(Glycine max[L.]Merr.)is an important crop that provides protein and vegetable oil for human consumption.As soybean is a photoperiod-sensitive crop,its cultivation and yield are limited by the photoperiodic conditions in the field.In contrast to other major crops,soybean has a special plant architecture and a special symbiotic nitrogen fixation system,representing two unique breeding directions.Thus,flowering time,plant architecture,and symbiotic nitrogen fixation are three critical or unique yielddetermining factors.This review summarizes the progress made in our understanding of these three critical yield-determining factors in soybean.Meanwhile,we propose potential research directions to increase soybean production,discuss the application of genomics and genomic-assisted breeding,and explore research directions to address future challenges,particularly those posed by global climate changes.

短期低温对大豆苗期生长和结瘤固氮的影响

采用早熟大豆品种“黑河43”、晚熟大豆品种“东农53”进行盆栽试验,出苗后进行连续7d的低温处理(LT),昼夜温度设定为13℃/3℃,以25℃/10℃为对照,从大豆出苗28d开始,每7d进行破坏性取样共4次,测定地上部和根干物质量、根瘤生物量、全氮含量及根瘤固氮量,研究苗期短期低温对大豆生长、结瘤和固氮能力的影响。结果表明:(1)大豆出苗后7d短期低温胁迫会延缓大豆生长发育。出苗后28~42d大豆生物量显著低于CK处理,出苗后49d低温胁迫对两个大豆品种生物量均无显著影响。(2)与CK相比,低温对“东农53”和“黑河43”根瘤形成的显著抑制作用分别出现在苗后42d和35d,根瘤数量分别降低了58.8%和72.0%。出苗后49d,低温使“东农53”和“黑河43”根瘤干重分别减少48.9%和48.5%。(3)低温刺激了两个品种大豆的生物固氮能力。在出苗后49d,低温处理“东农53”和“黑河43”的生物固氮量、单位根瘤生物量的固氮量和单个根瘤的固氮量分别增加了89.9%、118.9%,249.3%、172.6%,150.6%、114.2%。生物固氮百分率分别增加了26.4个和24.5个百分点。综合来看,大豆出苗后遭遇低温冷害会延缓生长发育,并抑制根瘤的形成与生长。低温胁迫解除后大豆生长逐步恢复。短期低温对大豆根瘤固氮量和根瘤固氮能力有刺激作用,对早熟品种“黑河43”刺激作用更明显。为了更好地适应东北大豆产区早春低温环境,推荐选用早熟品种大豆。

缺磷对已结瘤大豆生长和固氮功能的影响

磷对大豆生长起着非常重要的作用,然而区别磷在大豆生长和根瘤功能中的影响非常困难。为阐明根瘤形成后缺磷对大豆植株生长和固氮功能的影响,采用水培方法,营养液中供给正常磷酸盐浓度(30μmolL?1)条件下,待根瘤形成后,将营养液中的磷水平分别转换为0μmolL?1(无磷)和4μmolL?1(缺磷),研究缺磷对大豆生长和结瘤固氮的影响。结果表明,大豆根瘤具有固氮功能后,进行营养液中不同浓度的磷酸盐处理,从处理第9天开始,缺磷对大豆生物量的影响才表现出来,而缺磷对根瘤形成和生长的影响从处理开始就比较明显。处理第9天,无磷、缺磷和正常磷处理根瘤数比处理第3天分别增加11.8%、15.4%和20.0%,单位植株豆血红蛋白含量和单位根瘤豆血红蛋白含量都随磷浓度增加而增加。该结果表明,在保证大豆形成一定数目的根瘤后,缺磷会明显影响根瘤生长和固氮能力。

利用^(15)N标记研究氮素水平对大豆根瘤生长的影响

为了明确氮肥对大豆根瘤生长的影响,以东农47为试验材料,采用15N标记和砂培的方法,进行了氮素营养水平对大豆根瘤生长及与根瘤固氮相关性的研究。结果表明,随外源氮水平增加,根瘤干重呈先增加后降低的变化趋势,当营养液氮浓度为50 mg/L时,有利于根瘤的生长;根瘤生长需要一定量"启动氮","启动氮"的作用维持到第3片复叶完全展开时(V3)对大豆根瘤的生长最有利;在盛荚期(R4)和鼓粒始期(R5)补充外源氮的供给抑制根瘤的生长。根瘤干重与植株中根瘤氮积累量、根瘤氮比例呈极显著正相关,根瘤干重与植株中氮积累量呈显著正相关。

不同施氮量及供氮方式对大豆根瘤生长及固氮的影响

采用框栽试验方法,研究一次供氮(指在总氮量一定的情况下以基肥的形式一次性施入,设3种总供氮量处理:75 kg/hm2、150 kg/hm2、225 kg/hm2)和分次供氮(指在总氮量一定的情况下,分别于播种前、苗期、花期和鼓粒期分4次平均施入,与一次供氮处理相同,设3种总供氮量处理)对大豆根瘤干重和数量、氮积累量、固氮酶活性及豆血红蛋白含量的影响。结果表明:一次性施氮方式下,各处理地上氮积累量随着施氮量的增加而增加,根系氮积累量则随着施氮量的增加呈先增加后下降的趋势。分次施氮方式下,花期和鼓粒期根瘤干重和数量的变化趋势一致;苗期,各组织氮积累均以一次性施氮方式高于分次施氮和对照,且随着施氮量的增加而增加(地上);随着生育时期的推进,分次施氮对大豆植株氮积累促进作用增大,各组织氮积累表现为分次施氮>一次性施氮>对照。施氮量对固氮酶和豆血红蛋白含量的影响与一次性施氮方式下相似,但整体上较之抑制作用更强。

大豆保护性施氮的研究Ⅰ．氢醌在大豆保护性施氮中的效应

本文主要研究在施用尿素条件下，配加氢醌（ＨＱ）来延缓尿素的集中水解，以减少氨挥发并抑制其硝化作用，从而有效地缓解尿素对大豆共生固氮体系产生的严重抑制效应。结果表明，在一定的浓度范围内，氢醌对纯培养大豆根瘤菌生长。幼苗生长和初生结瘤无不利影响；对离体活性根瘤的呼吸活性和琥珀酸脱氢酶活性具有显著促进。在盆栽或田间条件下，尿素配加ＨＱ施用，同无ＨＱ等量尿素比较，显著提高了大豆结瘤量和单株固氮总活性（ＴＮＡ）；提高了大豆木质部中酰脲含量及其相对丰度，协调了大豆对根瘤氮和土壤（肥料）氮需求的矛盾，从而获得了显著的增产效果。

大气CO_2浓度升高对大豆根瘤量及其固氮活性的影响

通过开顶式气室控制CO2浓度，对盆栽大豆进行试验测定，研究了大气CO2浓度升高对大豆根系，重点是根瘤量和根瘤活性的影响。结果表明，大气CO2浓度升高，促进了大豆根系生长，大豆根体积、主根长、根鲜重均呈增长趋势，根冠比增加。CO2浓度为450、550、650和750μmol／mol时，与大气CO2背景浓度相比，在初花期，大豆根瘤数分别增加6．1％、15．9％、19．2％和26．5％，其中主根根瘤数增加较为显著，至鼓粒期，根瘤数增加幅度为7．8％～48．0％，增幅较初花期大，其中侧根根瘤量增加更多。同时，高CO2水平下，根瘤鲜重的变化与根瘤数基本一致。4种高CO2浓度下，初花期根瘤比固氮活性提高10．1％～24．0％，大于鼓粒期的6．0％～13．4％的增加幅度；单株根瘤固氮活性初花期增加10．6％～55．7％，鼓粒期则提高了20．0％～73．9％。

不同大豆品种根瘤固氮酶活性与固氮量差异研究

大豆根瘤固氮酶活性与固氮量是衡量大豆固氮能力的重要指标,也是合理施用氮肥的依据。试验采用框栽方法,应用15N示踪技术,研究了不同大豆品种根瘤固氮酶活性与固氮量差异。结果表明:4个供试大豆品种根瘤固氮酶活性、根瘤干重及根瘤固氮潜力都呈现先升后降的变化趋势,但品种间有显著差异;生育期短的大豆品种根瘤固氮酶活性高于生育期长的品种,根瘤干重则相反,生育期越长,根瘤干重越大;根瘤固氮潜力随大豆品种生育期的延长而增加。大豆植株氮素构成品种间存在明显的差异性,随着品种生育期的延长,根瘤固氮所占的比例提高;在大豆品种黑河41氮素构成中,土壤氮和根瘤固氮所占比例相同,其他品种均以根瘤固氮为主,其次是土壤氮,肥料氮所占的比例很低。

不同轮作方式对大豆孢囊线虫及大豆固氮能力的影响

１９９３～１９９４年的田间试验表明，不同轮作方式能显著地影响大豆根区孢囊线虫孢囊数量，每百克干土中孢囊数顺序为：重茬＞迎茬＞正茬．与正茬相比，重茬、迎再茬别降低单株根瘤重量２４．００％和３１．４８％；分别降低单株固氮能力３２．９４％和２２．５８％．而对根区根瘤菌数量及根瘤固氮酶活性没有明显的影响．

大豆光合生理生态的研究——第15报 大豆不同品种光合性状和固氮能力的比较

对大豆20个品种的光合性状和固氮能力研究表明,大多数新品种或高产品种植株中上部叶片较老品种具有较多的栅栏细胞层数和细胞及叶绿体数目;较大的叶厚度和比叶重;在生殖生长期具有较高的光合速率。同时大多数新品种具有较高的根瘤固氮酶活性和较大的幼茎段酰脲含量。大豆叶片光合速率和根瘤固氮酶活性及幼茎和荚皮酰脲含量均呈显著正相关。新品种与老品种相比具有较高的光合和固氮能力。

大豆不同生长阶段施用氮肥对生长、结瘤及产量的影响

以两种不同基因型的大豆品种为试材，研究不同阶段施用氮肥对大豆结瘤、生长及产量的影响。每公顷仅施２５ｋｇ的启动氮，两个大豆品种虽获得较高的根瘤干重，但仅靠其自身固氮功能，不能满足大豆生长及丰产的要求。在大豆开花期（Ｒ１）或种子形成始期（Ｒ５），每公顷再追施５０ｋｇ氮肥，两个大豆品种都获得了最高的干物质重和产量。在播种前或开花前期，施用过多的氮肥，严重抑制了两个大豆品种根瘤的生长，降低了其自身的生物固氮功能。

羊奶果的共生固氮及氮素的运转形式

对羊奶果根瘤的固氮、吸氢、防氧保护及固氮产物的运转进行研究。羊奶果是热带植物 ,离体根瘤固氮活性在 2 5～ 30℃时最高 ,且在离体 3h以内无显著变化 ;35～ 4 0℃的高温对根瘤吸氢活性有促进作用 ,离体根瘤的吸氢活性可保持 6 h以上 ;低温对固氮活性有明显的抑制作用。羊奶果为酰脲型植物 ,根、根瘤和木质部中都含有大量的尿囊酸 ,酰脲含量均占氮化合物总量的 5 0 %以上 ,说明它的根瘤和根都能将氮素转化成酰脲并经过木质部向上转运 ,还有部分氮素通过以丝氨酸、蛋氨酸为主的氨基酸形式运输。根瘤中未检测到血红蛋白 ,但有较高含量的 SOD、 ASA和

大豆结瘤固氮对磷素的需求

在温室条件下,利用水培技术,研究了磷对大豆生长、根生物量和根瘤固氮的影响。结果表明,大豆在生长初期表现出缺氮症状,随着磷浓度的增加,根瘤数、根瘤干重、氮和磷累积量均显著增加。在移栽后0d～14d,磷对大豆根瘤的生长影响不大,而在21d后。磷对根瘤生长的影响逐渐增大,移栽后35d,高磷处理比低磷增长了138％。植株不同部位磷含量大小顺序是：根瘤＞根＞地上部分。图5,表1,参6。

施氮对结瘤和非结瘤近等位基因大豆生长和固氮性状的影响

以结瘤和非结瘤近等位基因大豆品系为材料,利用砂培方式,分析了不同供氮水平对大豆生长、碳氮代谢、结瘤和固氮的影响。结果表明:供氮水平直接影响大豆生物量,结瘤大豆在N2水平(54 mg尿素·kg-1砂)时生物量达到最大(9.16 g·株-1);非结瘤大豆在N4水平(216 mg尿素·kg-1砂)时生物量达到最大(6.36 g·株-1)。植株体内碳氮含量与生物量的变化趋势相同。此外,结瘤品系N2水平大豆根瘤数最多(205个.株-1),极显著高于除N4以外的其它处理(P<0.01),该水平的共生固氮量最大,为275.92 mg·株-1,且与N3、N4、N5处理间差异极显著(P<0.01)。因此,结瘤和非结瘤大豆对外源氮的需求存在显著差异,该试验条件下的氮阻遏阈值为N2水平,即54 mg尿素·kg-1砂。

减氮对甜玉米//大豆间作系统大豆结瘤固氮特性的影响

为了探讨减氮和甜玉米//大豆间作对大豆结瘤和固氮特性的影响,通过大田定位试验(2015年春-2018年秋,共8季)对比了三种施氮水平:不施氮(N0,0 kg·hm^(-2))、减量施氮(N1,300 kg·hm^(-2))、常规施氮(N2,360 kg·hm^(-2)),两种间作模式:甜玉米//大豆2∶3间作(S2B3)、甜玉米//大豆2∶4间作(S2B4),以及不施肥单作大豆(SB)对大豆结瘤数、根瘤干重、固氮效率和固氮量及其稳定性的影响。结果表明:1)大豆根瘤数、根瘤干重、固氮效率和固氮量随着年季变化呈现明显的动态变化,春季大豆根瘤数和根瘤干重显著高于秋季,但秋季固氮效率和固氮量显著高于春季。2)施氮水平与种植模式极显著影响大豆固氮效率和固氮量,不施肥处理大豆固氮效率为S2B4(69.87%)>S2B3(60.64%)、SB(56.3%),但生物固氮量为SB(142.31 kg·hm^(-2))>S2B4(109.50 kg·hm^(-2))>S2B3(86.12 kg·hm^(-2))。3)间作甜玉米显著提高了大豆的固氮效率且随大豆种植比例的增加而增加,S2B4-N0、N1及N2的固氮效率分别比S2B3-N0、N1及N2高9.47%、3.41%、1.83%,但是,相同施氮水平下不同间作模式之间均无差异。4)减氮和间作甜玉米可显著提高大豆固氮率和固氮量的稳定性。总之,减氮和间作甜玉米均能促进大豆结瘤、提高生物固氮量及固氮效率。