大豆根瘤菌接种效应及竞争结瘤能力分析

为了筛选获得竞争结瘤能力强且对大豆生长发育、产量有积极影响的大豆根瘤菌菌株,以前期分离、鉴定、纯化的10株根瘤菌菌株为材料,对高匹配性大豆品种进行了田间接种试验。于大豆盛花期测定根瘤菌的结瘤数量及固氮酶性能。同时,对接种供试菌株及分离获得的根瘤菌菌株进行BOX-PCR并比较不同菌株之间的BOX分子指纹图谱,以此获得供试菌株的田间占瘤率。于成熟期测定大豆的主要生育特征、产量构成因子。结果表明:大豆接种根瘤菌后在整个生长期,叶片表现为颜色深绿,质地鲜嫩;根瘤菌可以显著促进大豆根系结瘤,增加单株根瘤数目。接种处理I4(即接种菌株112-1)的单株根瘤数最多,比不接种对照处理I0多32.95%,差异显著。接种根瘤菌处理的根瘤干重均显著低于不接种对照处理。其中,接种处理I4的根瘤干重最高。接种处理I6的固氮酶活性最高,与接种处理I4之间差异不显著;接种菌株的占瘤率为10%~90%,占瘤率变化幅度较大,平均占瘤率为50%。I4处理占瘤率最高为90%,I6处理(即接种菌株113-1)的占瘤率次之,为75%;菌株占瘤率与单株根瘤数呈极显著的正相关(r=0.82**),单株瘤干重与菌株占瘤率、单株根瘤数、固氮酶活性均呈现负相关(r=-0.387,r=-0.50,r=-0.13);I6处理株高最高,I4处理次之,两处理之间差异并不显著;I2、I4、I6处理的单株荚数、单株粒数相对较高,3处理之间差异并不显著;I4处理的大豆百粒重均显著高于其它菌液处理。根据接种根瘤菌对大豆结瘤固氮性能、产量等方面的影响,结合各根瘤菌株的竞争结瘤能力,筛选获得适于黑龙江哈尔滨地区大豆生产有广阔利用价值的高效大豆根瘤菌株112-1和113-1。     

接种根瘤菌对南疆春大豆结瘤和生长的影响

为筛选南疆结瘤能力强且对春大豆生长发育、产量有积极影响的大豆根瘤菌菌株,探讨大豆品种和根瘤菌的匹配性,以前期分离、鉴定、纯化的3株根瘤菌菌株为材料,对南疆地区3个春大豆品种进行了田间接种试验,测定大豆根瘤数、根瘤干重、大豆地上部干物质积累分配、产量及其构成因素的变化.结果表明:接种不同根瘤菌均能促进南疆不同春大豆品种根系结瘤,促进结瘤效果存在差异,黑农61接种T6能显著增加中后期根瘤数、根瘤干重,新大豆8号和石大豆2号接种T6和SN7-2能显著增加结瘤数和根瘤干重,SN7-2在春大豆生育前期、T6在生育后期作用明显;黑农61接种SMH12,新大豆8号和石大豆2号接种T6和SN7-2能显著促进干物质积累;新大豆8号接种SN7-2、石大豆2号接种T6、黑农61接种SMH12能促进干物质向生殖器官分配.接种根瘤菌能通过增加主茎节数、单株荚数和百粒重提高春大豆产量,新大豆8号与SN7-2、石大豆2号与SN7-2、黑农61与SMH12匹配性最好.     

中国和苏联大豆根瘤菌田间结瘤,固氮和接种效果的联合试验

黑龙江省与苏联远东地区间穆尔州邻近,生态条件相似。1989～1990年由黑龙江省农业科学院土肥所提供大豆根瘤菌菌株8852和8752,苏联全俄大豆所提供大豆根瘤菌菌株ЗД—4和ТО—13,分别在各自地区进行了大豆接种根瘤菌的联合试验。 一、试验方法 田间试验在黑龙江省农业科学院土肥所和黑河所进行。4个菌株每个为一个处理,加不接种为对照。每处理6垅、5米长、3次重复,随机区组设计。省农科院土肥所试验地为南部黑土,大豆品种为黑农33,     

外源类黄酮对大豆根瘤菌趋化性与结瘤效果的影响

以慢生型大豆根瘤菌(Bradyrhizobium japonicum)2-39为供试菌株,分别以不同浓度(5,10,15,20,25μg·mL~(-1))的大豆黄素、4-甲基伞形酮、金雀异黄酮、芒柄黄花素为诱导物,利用毛细管培养法对趋化的根瘤菌进行定量测定,研究不同种类和不同浓度的外源类黄酮对大豆根瘤菌趋化性的影响。选取对根瘤菌趋化性影响显著的外源类黄酮与大豆嫩丰16进行盆栽试验,进一步探究外源类黄酮对大豆结瘤效果的影响。结果表明:4种外源性类黄酮中,大豆黄素和金雀异黄酮对根瘤菌的趋化作用显著强于其它试验组,其最适作用浓度为15μg·mL~(-1)。盆栽试验结果显示:外源类黄酮大豆黄素和金雀异黄酮同时加入能够有效促进大豆结瘤。     

生态环境对根瘤菌竞争结瘤影响的研究进展

影响根瘤菌竞争结瘤能力的生态因素有土壤条件,肥力水平,土著根瘤菌的数量和类型,其它微生物,根瘤菌的接种方式和时间等.在土壤条件方面研究较多的是土壤类型、pH值.肥力水平主要指氮水平和有机质含量.     

采用gfp和rfp基因标记评价大豆根瘤菌竞争结瘤能力

采用二亲本接合方法,将绿色荧光蛋白基因(gfp)或红色荧光蛋白基因(rfp)分别导入与大豆品种中黄13相匹配的快生根瘤菌Sinorhizobium fredii SR25a、S.fredii USDA205、与慢生根瘤菌Bradyrhizobium japonicum 4222、B.japonicum 4230和B.japonicum 4534菌株中,得到6株标记成功的菌株,并检验标记菌株的外源基因在培养条件下和共生条件下的稳定性,以及对菌株结瘤性能的影响.进一步将带有不同荧光蛋白基因的供试菌株按等密度等体积配成9组,通过蛭石盆栽试验评价菌株的竞争结瘤能力.结果表明:外源基因能够遗传稳定,且不影响共生结瘤固氮行为,该标记方法可用于评价菌株竞争结瘤能力;慢生大豆根瘤菌的竞争能力明显高于快生大豆根瘤菌,其中慢生大豆根瘤菌B.japonicum 4534竞争能力最强,其占瘤率比慢生菌株B.japonicum 4230和B.japonicum 4222分别高出35%和90%.结果证明gfp和rfp双标记技术为根瘤菌竞争结瘤能力评价提供了直观、简便、准确的检测手段.     

氮肥与根瘤菌配施对南疆春大豆结瘤和生长的影响

为了探讨最适合南疆地区的氮肥和根瘤菌配施方案,设置3个施氮水平(N0:0 kg·hm-2;N1:90 kg·hm-2;N2:180 kg·hm-2)与3种根瘤菌(SMH12、T6和SN7-2)拌种的田间试验,研究主栽大豆品种新大豆8号结瘤情况、干物质积累及分配、产量及其构成因素对施氮及接种根瘤菌的响应.结果表明:随着施...     

耐旱大豆根瘤菌的筛选及其接种效应

采用聚乙二醇(PEG)6000模拟干旱的试验方法,从分离自黄淮海地区的19株根瘤菌和2株参比菌株中初步筛选得到能够在该条件下生长良好的根瘤菌7株。进一步进行土壤盆栽干旱胁迫复筛试验,通过根瘤数量、大豆植株干重和含氮量等指标分析,获得了3株耐旱根瘤菌株B.japonicum 4788、B.japonicum 4792和B.japonicum USDA110;测定了在盆栽条件下接种B.japonicum 4792大豆植株耐旱性相关生理的指标,与未接种对照相比,接种根瘤菌的大豆叶片中甜菜碱、SOD酶和叶绿素的含量都有不同程度的增加,丙二醛的含量减少,表明接种根瘤菌可以提高大豆的耐旱性能。     

磷和根瘤菌交互作用对大豆结瘤和生长的影响

为了阐明营养液中磷浓度和不同数量根瘤菌的交互作用对大豆结瘤和生长的影响,在营养液培养条件下,进行3种磷水平[P1-0、P2-4和P3-30(μmol P·I-1)]和3种根瘤菌浓度[RI-102、R2-103.5和R3-105(CFU·mL-1)]处理,培养大豆25 d,测定大豆生物量和根瘤性状指标.结果表明:在相同根瘤菌数量条件下,不同磷浓度对大豆总生物量的影响不大,对根冠比的影响较大营养液中接种的根瘤菌浓度越高,根瘤原基数量越夫,低磷明显抑制根瘤原基发育形成根瘤,进而减少了低磷处理大豆根瘤数量从P1到P3,R1恨瘤菌浓度处理根瘤原基形成根瘤的几率分别为:79%、86%和100%.营养液中磷浓度促进生物量向根瘤中分配,高磷处理相对根瘤生物量较高随着生育进程,大豆各组织器官含磷量逐渐降低,在所有取样时期含磷量均表现为根瘤>根>地上部;含氮量表现为根瘤>地上部>根.可见磷和根瘤菌对大豆生长和结瘤形成有交互作用.     

根瘤菌和复合促生菌对大豆结瘤和生长的影响

为了研究不同根瘤菌和复合促生菌单独使用及复合后对大豆结瘤和生长的影响。采用大豆单独接种2种快生大豆根瘤菌、2种慢生大豆根瘤菌及根瘤菌分别与复合促生菌交叉接种,再种植的方法,观察大豆结瘤与生长情况。结果表明:(1)大豆结荚初期大豆快生和慢生根瘤菌处理的大豆地上部鲜重、地下部鲜重、结荚数、豆荚鲜重等生物学性状显著优于其它处理;快生根瘤菌处理的大豆结瘤率显著高于其它处理,慢生根瘤菌处理的总根瘤数和根瘤重量显著高于其它处理;(2)大豆结荚后期,对照处理的地上部鲜重、根鲜重显著高于其它处理,慢生根瘤菌和复合促生菌处理的根瘤数显著高于其它处理,快生根瘤菌剂处理的大豆粗蛋白含量显著高于其它处理,而复合促生菌处理的大豆产量显著高于其它处理。虽然快生根瘤菌两处理有利于地上部、地下部、有效根瘤数、土壤碱解氮的提升,但复合促生菌处理的大豆产量、土壤速效磷和土壤有效钾都显著高于其它处理,因此表明复合促生菌处理能增加大豆产量。     

基于大豆导入系群体结瘤相关性状QTL定位及基因挖掘

氮元素是植物生长所需的重要营养元素之一,生物固氮是大豆生长所需氮元素的重要来源。本实验室以绥农14为母本,野生豆ZYD00006为父本构建一套覆盖野生大豆全基因组的导入系群体。基于大豆导入系群体对结瘤数目和干重进行QTL定位,定位到3个QTL与根瘤干重相关,分布在N、M和D2这3个连锁群上,定位到5个QTL与根瘤数相关,分布在K、F、J和D2这4个连锁群上,在D2连锁群这两个性状有重叠区段(7.20-7.79Mb)。针对这个重叠区段的63个基因进行基因注释,选择到6个与共生、抗病相关的基因作为候选基因进行下一步验证。qRT-PCR分析表明根瘤菌侵染期间Glyma.17G097000基因表达模式与对照相比差异很大。Glyma.17G097000属于GmHIR基因家族,在大豆基因组中发现了11个家族成员。GmHIR家族基因结构相似性很高,基因表达有组织特异性。大豆HIR蛋白有Stomatins和Prohibitin两个结构域,能够参与离子通道调节等生理过程,与植物抗病和细胞周期有关。GmHIR基因来源于四个祖先,进化过程中是高度保守的。对GmHIR基因家族11个成员qRT-PCR检测,结果显示根瘤菌感染期间Glyma.05G029800、Glyma.09G154400和Glyma.17G097000这三个基因与对照相比表达模式差异较大。结果表明这三个基因可能在大豆共生体系建立中起着重要的作用,参与根瘤菌与大豆共生体系建立过程中的离子通道调节和免疫反应。本研究为大豆-根瘤菌共生机制研究奠定基础并提供有效候选基因。     

连作对大豆根瘤形成和固氮功能影响的研究展望

通过分析大豆连作障碍因子对大豆生长和品质的影响,阐明目前连作障碍因素对大豆影响研究中存在的问题,提出研究连作障碍因素对大豆根瘤形成、生长发育和固氮功能的影响是未来大豆连作障碍研究中亟待解决的问题,并指明研究的具体内容和拟要达到研究目标.     

荫蔽对大豆主要性状的影响及大豆耐荫性鉴定方法研究初报

模拟间套种的荫蔽条件，设置不同的遮光处理，研究荫蔽对大豆13个主要性状的影响。结果表明，结荚期株高、收获期株高、最低结荚高度、结荚期生物产量、单株荚数、单株粒数、单株产量和百粒重等8个性状发生显著变异，变异与荫蔽程度极显著相关，其中，前3者为正相关，其余5性状为负相关；结荚节位和主茎直径变异极显著，分枝数变异显著，但与荫蔽程度相关性不显著；主茎节数和单株瘪荚数变异不显著。选择受荫蔽影响显著且与荫蔽相关性显著的株高、最低结荚高度、结荚期生物产量、单株荚数、单株粒数、单株粒重和百粒重等7个性状为指标性状，以综合耐荫系数为指标，采用综合方法评定大豆种质的耐荫性，将耐荫性鉴定级别划分为五级。     

An improved method for rapid quantitative analysis of the insoluble dietary fiber in common cereals and some sorts of beans

An improved rapid analysis for determining the content of insoluble dietary fiber (IDF) in common cereals and some sorts of beans is described in this paper. The procedure includes starch gelatinization in water bath for 20 min at 100 °C and 2.5% (w/w) α-amylase hydrolyzed reaction followed by neutral detergent wash and acetone extraction. Compared with 1.5 h for filtration (estimated) and 18 h for the enzymatic hydrolysis required by the typical American Association of Cereal Chemists (AACC) method, the filtration and enzymatic treatment procedures in the improved method was completed within 15 min and 1.5 h, respectively. The length of time for the filtration and the enzymatic hydrolysis for the improved method was significantly shortened from 19.5 h (AACC method) to 1.75 h. In addition, orthogonal array design (OAD) has been applied to optimize parameters of the improved method. The recovery yield of microcrystalline cellulose was 97.75% (w/w), in agreement with the result obtained using the typical AACC method, demonstrating the reliability of the improved method. Furthermore, several common cereals and beans were employed to validate the accuracy and universality of this improved method.     

Cereal protein analysis via Dumas method: Standardization of a micro-method using the EuroVector Elemental Analyser

The Dumas method (DM) of protein analysis in foods was standardized using two cereal references in order to identify the main sources of error during analysis; specifically, the study sought to establish which of the apparent critical factors (ACFs) were really significant influencing factors (IF) for the application of the DM to protein analysis in cereals.     

Stability analysis of seven agronomic traits for soybean [(Glycine max (L.) Merr.] Tokachi nagaha and its derived cultivars using the AMMI model

The stabilities of seven agronomic traits were analyzed and the general stabilities of soybean [Glycine max (L.) Merr.] accessions were evaluated based on the additive main effects and multiplicative interactions (AMMI) model using the founder parent Tokachi nagaha and 137 of its derived cultivars as materials. The objective was to provide a theoretical basis for effectively using germplasm in soybean breeding and production. Analysis of variance (ANOVA) showed that genotype, environment, and genotype by environment interactions were significantly different for each trait. The first three interaction principal components axes (IPCA) were highly significant, accounting for 61.28–70.00% of the total variation. The stability differed for the different traits. 50 cultivars with high general stabilities were identified. The general stability of Tokachi nagaha was moderate, as the stability coefficients (D_i) of its seven traits were relatively high; this must be considered by breeders using this cultivar as a breeding parent. There were significant positive correlations between the phenotypic values and their own D_i values for number of branches per plant, number of pods per plant, number of seeds per plant, and seed weight per plant. This would lead to the expectation that the phenotypic stability would be lower when a cultivar had more branches, pods, seeds, and high per plant yields. Thus, it appears difficult to breed cultivars that simultaneously have high yields and high stability.     

Integrating models of relative abundance of species with the dry-weight-rank method for the botanical analysis of forest understorey vegetation

The objective of this work was to determine the applicability of the dry‐weight‐rank (DWR) method for evaluating the botanical composition of forest understorey vegetation. An analysis of plant species abundance was carried out, and instead of ranking the three most abundant species, as is commonly used, up to twelve ranks were scored. Concurrently, four models of relative abundance distribution (RAD) of species were compared for their ability to explain the abundance of species in the study area. The Power‐fraction model resulted in the best goodness‐of‐fit and it was subsequently used to produce the coefficients for the DWR method. Lin's concordance correlation coefficient, the adjusted coefficient of determination, the residual standard deviation and Spearman's rank‐order correlation coefficient indicated a good performance of the DWR method. Biomass data and the Shannon index for diversity were also considered. Further analyses showed that there was a trade‐off between the number of ranks scored and the accuracy of the botanical composition produced by the DWR method. It is concluded that, so long as the RAD model that explains the distribution of plant species is known, the DWR method can be applied to forest understorey vegetation.     

A simple method for squeezing juice from rice stems and its use in the high-throughput analysis of sugar content in rice stems

Sugar content in rice (Oryza sativa L.) stem is an agronomically important trait for rice used in straw silage or whole-crop silage. However, the mechanisms underlying sugar accumulation in rice stems remain unclear, mainly due to the time-consuming method for measuring sugar content. Here, we established a simple method for squeezing stem juice from rice plants, similar to that used during breeding selection in sugarcane or sorghum. The Brix value of the stem juice, which can easily be measured using a portable refractometer, significantly correlated with the soluble sugar contents in the stem juice and tended to correlate with those in stem tissues. This indicates that the Brix value of the stem juice can be used for estimating the sugar content in rice stems. This simple estimation method will be a useful tool for high-throughput analysis of sugar content in rice stems during mutant screening, QTL analysis, and breeding selection.     

An analysis of the hybrid nature ofsolanum xedinense Berth, using molecular, cytological and crossability studies

Summary Solanum xedinense Berth., a pentaploid (2n=5x=60) interspecific hybrid between the cultivated potato and the wild speciesSolanum demissum Lindl., represents evidence of genetic exchange between wild and cultivated species. It has been found in Central Mexico at altitudes between 2000 and 3500 meters, growing in and along the edges of cultivated fields, roadside thickets, forest margins and along irrigation ditches. This study examined the hybrid nature of S.xedinense using molecular, cytological and crossability data. RAPD and AFLP markers were used to supportS. xedinense as a hybrid between the cultivated potato andS. demissum. Meiotic analysis and crossability studies were used to examine the potential ofS. xedinense to form offspring. Several accessions ofS. tuberosum L. subsp.tuberosum and subsp.andigena Hawkes,S. demissum and an accession ofS. xedinense were used in the study. Artificial hybrids betweenS. tuberosum subsp.tuberosum andS. demissum were included for comparison. Molecular analysis positioned the natural and artificial hybrids as an intermediate group between the parental populations, supporting their relationship with the parental populations. Meiosis was abnormal in both natural and artificial hybrids, and reduced crossability was noted forS. xedinense. Although gene flow appears to happen between wild and cultivated potato, the potential for the natural hybrids to become established may depend mostly on their ability to clonally propagate themselves.     

A simple and rapid procedure for the analysis of reducing,total and individual sugars in potatoes

Summary A simple analytical scheme based on the micro-colorimetric reducing sugar methods of Somogi is described which permits the reducing and total sugars, as well as glucose, fructose and sucrose, to be determined in aqueous methanolic extracts of potatoes. The reducing and total sugars reskpectively-and thus, by difference, sucrose — are determined by estimating reducing power before and after inversion with hot dilute HCl. The individual reducing sugars are estimated from the decrease in reducing power after complete removal of glucose with glucose oxidase. The procedure is rapid and convenient and permits estimations to be made to within approximately ±5% of actual values.

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Zusammenfassung Es wird ein einfaches analytisches Verfahren beschrieben, das es erlaubt in w?ssrigen Methanolextrakten die reduzierenden Zucker und den Gesamtzucker sowohl als auch Glukose. Fruktose und Saccharose einzeln zu bestimmen. Aliquote Teile des Extraktes werden in geeichte Reagenzgl?ser gebracht und das Methanol im kochenden Wasserbad abgedampft. Bei einigen Proben wird die Saccharose mit heisser 0.1M HCl (5 Min) gespalten und die L?sung anschliessend mit Natriumkarbonat neutralisiert. Einige Proben werden mit gereinigter Glukoseoxidase behandelt, um die Glukose quantitativ zu entfernen. Die reduzierenden Zucker und die Gesamtzuckermenge werden dann durch die Zunahme des Reduktionsverm?gens vor und nach der Inversion bestimmt, durch Anwendung der empfindlichen und spezifischen mikrokolorimetrischen Kupferreduktionsmethode von Somogi (1952). Die Differenz ergibt den Gehalt an Saccharose. Die einzelnen reduzierenden Zucker werden durch Abnahme der Reduktionskraft nach der Behandlung der Probe mit Glukoseoxidase bestimmt. Das gesamte Verfahren ist schnell und einfach und bringt ausgezeichnete Wiedergewinnungswerte sowohl mit einfachen Zuckergemischen als auch mit Kartoffelextrakten, denen bekannte Mengen von Zuckern zugesetzt wurden. Im allgemeinen lagen die Wiedergewinnungswerte innerhalb von ±5% der erwarteten Werte (Tabelle 1). Tabelle 1 zeigt, dass die Analysenergebnisse für den Gesamtzuckergehalt in Kartoffelextrakten. die nach der beschriebenen Methode erhalten wurden, gut mit solchen übereinstimmten, die mit v?llig unabh?ngigen Methoden wie z.B. der Anthronreaktion erreicht werden.

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Résumé Une méthode d'analyse simple est décrite, permettant la détermination des sucres totaux, des sucres réducteurs et individuellement du glucose, du fructose et du saccharose contenus dans des extraits alcooliques de pommes de terre (méthanol à 85%). Après évaporation au bain marie du méthanol d'une fraction aliquote des extraits placés dans des séries de tubes calibrés. I'inversion du saccharose de quelques-uns des échantillons est effectuéc par 0.1M HCl chaud, pendant 5 minutes; puis les solutions sont neutralisées avec du carbonate de sodium. Le glucose est entièrement éliminé des autres échantillons par un traitement à la glucoseoxydase purifiée. Les sucres réducteurs. Les sucres totaux-et désormais, par différence, le saccharose —sont alors évalués par l'augmentation du pouvoir réducteur après inversion. à l'aide d'un microcolorimétre sensible et spécifique de la réaction de réduction du cuivre selon la méthode de Somogi (1952). Chaque sucre réducteur est déterminé par la diminution du pouvoir réducteur de la solution. résultant du traitement à la glucose-oxydase. Le procédé est rapide et commode; il a donné d'excellents recoupements aussibien avec les tests effectués sur des mélanges de sucres en solution que sur des extraits de pommes de terre auxquels on a rajouté des quantités déterminées de sucres. En général, ces recoupements ont été de ±5% des valeurs conneus (tableau 1). Les résultats obtenus par l'analyses des sucres totaux dans les extraits de pomme de terre en utilisant la méthode décrite, sont en concordance avec ceux obtenus par une autre méthode basée sur la réaction à l'anthrone (tableau 1)..