低分子量有机酸对红壤无机态磷转化及酸度的影响

以鄂南、赣北两红壤样品为材料,加入不同有机酸并经室温培养后,测定不同P组分、pH及活性Al含量的变化。结果表明,供试有机酸均使土壤Ca2－P含量增高,增幅大小依次为柠檬酸＞苹果酸＞琥珀酸＞乙酸;2种土壤的Ca8－P和Ca10－P含量无明显变化规律,Fe-P、Al-P和O－P含量有所下降,除乙酸处理的土壤pH值无显著变化外,其它有机酸的加入使pH下降0．65－1．96;有机酸引起活性Al量增多,除乙酸处理的变化较小外,其它有机酸或混合物的加入使土壤中0．02mol.L^-1CaCl2提取Al增加4．7－50．3倍,1mol.L^-1提取Al增加4．0－67．3倍。可见,有机酸具有双重作用,既增加P的有效性,又增加土壤酸度和Al毒。     

有机酸对土壤无机态磷转化和速效磷的影响

土壤有效磷含量低是影响作物生产的重要限制因素之一.作物根分泌活化难溶性磷的有机酸对改善其磷素营养具有重要意义.采用张守敬和Jackson无机磷分级方法,以湖北省3种pH值土壤为材料,加入不同磷源和有机酸,经过室温培养后,测定速效磷含量和无机磷组分的变化.结果表明:施磷显著提高了土壤中速效磷含量,中性土、酸性土Fe-P和Al-P含量大幅上升, Fe-P占增加量的50%以上,而碱性土Ca-P含量显著增加.加施有机酸使中性土速效磷含量增多,除苹果酸处理的变幅较小外,草酸和柠檬酸的加入速效磷显著增加.由于有机酸的作用,中性土Al-P含量下降,Ca-P含量上升,变幅大小依次为草酸＞柠檬酸＞苹果酸;酸性土中Al-P含量呈下降趋势,碱性土中Ca-P含量有不同程度的减少,3种土壤中O-P含量均有所增加.说明有机酸活化的磷主要来源于中性土和酸性土Al-P、Fe-P及碱性土Ca-P中的磷,同时有机酸能够促进土壤中闭蓄态磷(O-P)的形成与积累.     

低磷和铝毒胁迫条件下菜豆有机酸的分泌与累积

以水培方式研究了低磷、铝毒胁迫条件下,不同菜豆基因型根系有机酸的分泌及其在植穆不同部位的累积,结果表明,低磷,铝毒胁迫诱导菜豆有机酸的分泌与累积存在显著的基因差异。低磷、铝毒胁迫诱导菜豆主要分泌柠檬酸、酒石酸和乙酸,其中,50μmol/LAl^3 诱导柠檬酸分泌量最高;低磷（小于20μmol/LH2PO4^－）胁迫诱导柠榨菜酸分泌量显著高于高磷处理,但低磷处理之间差异不明显,铝毒胁迫诱导菜豆有机酸的分泌与累积显著高于低磷胁迫处理,低磷,铝毒胁迫植株不同部位有机酸的含量为叶片大小根系,低磷,铝毒胁迫时,G842菜豆型柠檬酸有机酸分泌总量显著高于273、AFR和ZPV,其干重和磷吸收明明显于大G273,AFR和ZPV,且铝吸收量小于G273,AFR和ZPV,说明,G482菜豆基因型对低磷,铝毒的适应能力强于G273,AFR和ZPV基因型,菜豆有机酸,,尤其柠檬酸的分泌是其适应低磷、铝毒胁迫的重要生理反应。     

白浆土-植物系统营养物质转化机制及其有效性研究II.白浆土无机磷形态变化

采用室内培养和微区试验研究表明,施入白浆土中无机磷在短时间内（24h、8d）主要向Fe-P和Al-P转变,较长时间后（80d）,Al-P向Fe-P转化,Ca-P变化较弱,土壤中无机磷形态转化与白浆土本身的土壤属性具有密切关系。施用有机肥改变了土壤中无机磷形态的转化过程,其与土壤中的铁、铝氧化物反应形成的“Al-有机质-P”和“Fe-有机质-P”是作物较易利用的有效P源。     

低磷和干旱胁迫对大豆植株干物质积累及磷效率的影响

土壤缺磷和季节性干旱已经成为南方酸性红壤地区大豆生产的主要限制因素之一。选取2个大豆品种巴西10号(磷高效)和本地2号(磷低效),研究其在不同磷素(0,15, 30 mg/kg P)和水分处理(分别在开花期和结荚期进行干旱胁迫)下的反应,从植株生物量、叶绿素含量、磷效率指标等方面研究不同基因型大豆对水磷耦合胁迫的适应机制。研究结果表明,随着土壤磷素水平的增加,两个品种的生物量和叶片叶绿素含量显著增加,根冠比则显著下降。在同一磷素水平处理下,干旱胁迫则导致较高的根冠比,对叶片叶绿素含量影响不大,两个品种表现一致。两个基因型大豆受到干旱胁迫后,其产量均显著低于正常水分处理。中等施磷能显著提高两个大豆品种的产量,但高磷处理对产量的增加幅度有限,甚至高磷处理还造成本地2号减产。巴西10号的产量随土壤中磷素的增加而增加,而本地2号的产量则为中磷>高磷>低磷,不管是磷处理还是水分处理,巴西10号的产量均高于本地2号。无论是花期干旱还是结荚期干旱,巴西10号和本地2号的根磷效率比、磷吸收效率及磷转移效率均随土壤磷浓度的增加而增加,磷利用效率则降低。总体上来讲,巴西10号的磷吸收效率和利用效率高于本地2号,而根磷效率比、磷转移效率则小于本地2号。     

白浆土-植物系统营养物质转化机制及其有效性研究Ⅱ.白浆土无机磷形态变化

采用室内培养和微区试验研究表明,施入白浆土中无机磷在短时间内（２４ｈ、８ｄ）主要向ＦｅＰ和ＡｌＰ转变,较长时间后（８０ｄ）,ＡｌＰ向ＦｅＰ转化,ＣａＰ变化较弱,土壤中无机磷形态转化与白浆土本身的土壤属性具有密切关系．施用有机肥改变了土壤中无机磷形态的转化过程,其与土壤中的铁、铝氧化物反应形成的“Ａｌ有机质Ｐ”和“Ｆｅ有机质Ｐ”是作物较易利用的有效Ｐ源     

低磷对大豆主根伸长生长的影响

文章采用卷纸培养和分层琼脂培养的方法,研究磷对大豆主根伸长影响的结果表明:低磷[0.2 μmol(KH2PO4)·L-1]显著促进大豆主根伸长,特别是延长大豆主根根尖至最新侧根间的距离;组织切片表明,低磷对主根伸长的促进主要是通过延迟主根伸长区的分化实现的,并且低磷对主根的促进作用不受亚磷酸盐的影响.琼脂分层培养的结果表明,在磷分布不均匀的条件下,低磷影响主根的仲长生长,上层或下层不施磷的大豆主根伸长均有增加.     

缺磷条件下低分子量有机酸对大豆氮积累和结瘤固氮的影响

采用砂培方法在温室条件下研究了低分子量有机酸柠檬酸、草酸、苹果酸及3种酸的混合物对大豆植株氮素积累、结瘤和固氮的影响.结果表明：低分子量有机酸对大豆植株氮素积累有显著的抑制作用,使大豆地上部各时期氮素积累量的降低幅度分别为：苗期17.6%～44.9%,花期29.8%～88.4%,鼓粒期9.18%～69.6%,成熟期2.21%～41.7%;低分子量有机酸对大豆根瘤生长和固氮能力也有显著影响,表现为使根瘤数量、根瘤固氮酶活性和豆血红蛋白含量显著降低,降低幅度分别为11.4%～59.6%,80.5%～91.7%和11.9%～59.9%,从而使大豆的固氮效率降低,最终导致大豆的固氮量较对照显著降低(降低幅度9.71%～64.5%).低分子量有机酸对大豆氮积累、根瘤生长和固氮能力的抑制作用随浓度的增加而增加.3种有机酸中,草酸的抑制作用相对大于柠檬酸和苹果酸,3种有机酸混合后,抑制作用加强.     

外源有机酸对玉米磷吸收及其生长发育的影响

以玉米品种郑单958为材料,通过施加外源有机酸的盆栽试验方法,设置了土壤中不施磷肥和有机酸(CKo)、仅施磷肥(CKP)、施柠檬酸(Tca)、施苹果酸(Tma)和施草酸(Toa)处理,研究外源有机酸对玉米生长过程中的磷素吸收及其生长发育的影响.结果显示:(1)3种外源有机酸均可显著提高土壤中速效磷含量和磷素利用效率,Tca、Tma和Toa处理玉米根际土壤的速效磷含量分别为CKo的2.25、1.96和2.04倍,且各处理间的磷素利用效率依次为:CKp>Tca>Tma>Toa>CKo.(2)在拔节期、抽雄期和成熟期,Tca处理玉米的磷素利用效率依次为CKo的1.16、1.05和1.04倍.(3)3种外源有机酸均可显著提高玉米根系体积和根系活力、单株玉米的氮磷钾的累积量、玉米的生物量及相对籽粒产量.研究表明,外源柠檬酸、苹果酸和草酸均可显著提高玉米根际土壤中固态磷的溶解,促进植株体内营养元素的累积,保证植物生长发育的需要,从而提高玉米的生物量和产量;并以外源柠檬酸处理对玉米磷吸收及其生长发育效果最好.     

大豆种子储藏性磷对幼苗适应低磷胁迫的作用

选用种子大小不同、磷效率不同的两个大豆品种‘巴西10号’(B10)和‘本地2号’(L2),在不同供磷条件下进行营养液浇灌沙培,从大豆萌发至2片三出复叶完全展开期测定植株主要器官总磷、可溶性磷浓度、子叶可溶性蛋白、酸性磷酸酶比活性、植酸酶比活性的变化动态,探讨储藏性磷在大豆幼苗期适应磷胁迫中的作用。结果发现:(1)磷效率不同的两个大豆品种的种子中磷含量差异显著。(2)大豆萌发和幼苗生长过程中子叶的磷逐渐转入根、茎、叶中,并以转入叶中的磷最多,其中磷高效品种B10在发芽过程中子叶磷向各个器官转移的总磷量要高于磷低效品种L2,且持续时间长。(3)大豆萌发和幼苗生长过程中外源供磷水平显著影响子叶磷的转移,在外源供磷充足条件下各器官中总磷均高于低供磷条件,子叶中磷和外源磷存在补偿关系。(4)磷高效品种B10子叶中酸性磷酸酶活性在低磷条件下显著高于高磷条件,但磷低效品种L2在高、低磷间无显著差异。研究表明,大豆种子储藏性磷在幼苗期耐低磷能力建立方面具有重要作用。     

利用聚鸟氨酸提高大豆原生质体外源基因的转化效率（简报）

The foreign Bt gene was transferred into protoplasts of soybean using PEG and PLO methods, respectively. The result indicated that the transformation frequency of PLO method was about 0.1% higher than PEG method. The PCR and Southern blotting analysis of the regeneration plants confirmed the integration of foreign gene into the genome of soybean.     

Plant regeneration from protoplasts of soybean (Glycine max L.)

Protoplasts were isolated from immature cotyledons of six cultivars of Glycine max L. and cultured in the KP8 liquid medium supplemented with 0.2 mg/L 2,4-D, 1 mg/L NAA and 0.5 mg/L ZT. The protoplasts started to divide after 3–5 days of culture. Sustained divisions resulted in mass production of cell colonies and small calli in 6 weeks. The calli further grew to 2–3 mm on the gelritesolidified K8 medium and were transferred onto the MSB medium with 1 mg/L 2,4-D and 0.25 mg/L BA, to obtain compact and nodular calli. Shoot formation was initiated on MSB medium with 0.15 mg/L NAA, and BA, KT and ZT, 0.5 mg/L of each, with or without 500 mg/L CH. It was followed by plant regeneration. So far, 87 plants have been regenerated from 4 cultivars, and normal seeds were obtained from them after transplanting into pots.Abbreviation IAA indol-3-acetic acid - NAA naphthalene acetic acid - 2,4-D 2,4-dichlorophenoxy acetic acid - KT kinetin - BA 6-benzyladenine - ZT zeatin - CH casein hydrolysate     

Cadmium-induced senescence in nodules of soybean (Glycine max L.) plants

The relationship between cadmium-induced oxidative stress and nodule senescence in soybean was investigated at two different concentrations of cadmium ions (50 and 200 μM), in solution culture. High cadmium concentration (200 μM) resulted in oxidative stress, which was indicated by an increase in thiobarbituric acid reactive substances content and a decrease in leghemoglobin levels. Consequently, nitrogenase activity was decreased, and increases in iron and ferritin levels were obtained. Senescent parameters such as ethylene production, increased levels of ammonium and an increase in protease activity were simultaneously observed. Glutamate dehydrogenase activity was also increased. Peroxidase activity decreased at the higher cadmium concentration while the lower cadmium treatment produced changes in peroxidase isoforms, compared to control nodules. Ultrastructural investigation of the nodules showed alterations with a reduction of both bacteroids number per symbiosome and the effective area for N₂-fixation. These results strongly suggest that, at least at the higher concentration, cadmium induces nodule senescence in soybean plants.     

Molecular-genetic identification and certification of soybean (Glycine max L.) cultivars

An approach to certification of soybean genotypes has been developed. The procedure employs three methods of DNA analysis based on polymerase chain reaction (PCR): PCR with arbitrary primers (AP PCR), simple sequence repeat polymorphism (SSRP) analysis, and inter-simple sequence repeat (ISSR) analysis. The approach to certification proposed may be used in both genetic and breeding research and seed production. A "certificate" form that reflects the unique characteristics of each cultivar studied is proposed. The results of molecular genetic analysis of allele distribution in genotypes of soybean from different ecological geographic zones permit estimation of the adaptive significance of individual alleles.     

Host genetic control of symbiosis in soybean (Glycine max L.)

Genes controlling nitrogen-fixing symbioses of legumes with specialized bacteria known as rhizobia are presumably the products of many millions of years of evolution. Different adaptative solutions evolved in response to the challenge of survival in highly divergent complexes of symbionts. Whereas efficiency of nitrogen fixation appears to be controlled by quantitative inheritance, genes controlling nodulation are qualitatively inherited. Genes controlling nodulation include those for non-nodulation, those that restrict certain microsymbionts, and those conditioning hypernodulation, or supernodulation. Some genes are naturally occurring polymorphisms, while others were induced or were the result of spontaneous mutations. The geographic patterns of particular alleles indicate the role of coevolution in determining symbiont specificites and compatibilities. For example, the Rj4 allele occurs with higher frequency (over 50%) among the soybean (G. max) from Southeast Asia. DNA homology studies of strains of Bradyrhizobium that nodulate soybean indicated two groups so distinct as to warrant classification as two species. Strains producing rhizobitoxine-induced chlorosis occur only in Group II, now classified as B. elkanii. Unlike B. japonicum, B. elkanii strains are characterized by (1) the ability to nodulate the rj1 genotype, (2) the formation of nodule-like structures on peanut, (3) a relatively high degree of ex planta nitrogenase activity, (4) distinct extracellular polysaccharide composition, (5) distinct fatty acid composition, (6) distinct antibiotic resistance profiles, and (7) low DNA homology with B. japonicum. Analysis with soybean lines near isogenic for the Rj4 versus rj4 alleles indicated that the Rj4 allele excludes a high proportion of B. elkanii strains and certain strains of B. japonicum such as strain USDA62 and three serogroup 123 strains. These groups, relatively inefficient in nitrogen fixation with soybean, tend to predominate in soybean nodules from many US soils. The Rj4 allele, the most common allelic form in the wild species, has a positive value for the host plants in protecting them from nodulation by rhizobia poorly adapted for symbiosis.     

Sonication-assisted Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merrill] embryogenic suspension culture tissue

Successful transformation of plant tissue using Agrobacterium relies on several factors including bacterial infection, host recognition, and transformation competency of the target tissue. Although soybean [Glycine max (L.) Merrill] embryogenic suspension cultures have been transformed via particle bombardment, Agrobacterium-mediated transformation of this tissue has not been demonstrated. We report here transformation of embryogenic suspension cultures of soybean using “Sonication-Assisted Agrobacterium-mediated Transformation” (SAAT). For SAAT of suspension culture tissue, 10–20 embryogenic clumps (2–4 mm in diameter) were inoculated with 1 ml of diluted (OD_600nm 0.1–0.5) log phase Agrobacterium and sonicated for 0–300 s. After 2 days of co-culture in a maintenance medium containing 100 μM acetosyringone, the medium was removed and replaced with fresh maintenance medium containing 400 mg/l Timentin^?. Two weeks after SAAT, the tissue was placed in maintenance medium containing 20 mg/l hygromycin and 400 mg/l Timentin^?, and the medium was replenished every week thereafter. Transgenic clones were observed and isolated 6–8 weeks following SAAT. When SAAT was not used, hygromycin-resistant clones were not obtained. Southern hybridization analyses of transformed embryogenic tissue confirmed T-DNA integration. Received: 22 August 1997 / Revision received: 22 October 1997 / Accepted: 11 November 1997     

QTL analyses of seed weight during the development of soybean (Glycine max L. Merr.)

At harvest traits such as seed weight are the sum of development and responses to stresses over the growing season and particularly during the reproductive phase of growth. The aim here was to measure quantitative trait loci (QTL) underlying the seed weight from early development to drying post harvest. One hundred forty-three F(5) derived recombinant inbred lines (RILs) developed from the cross of soybean cultivars 'Charleston' and 'Dongnong 594' were used for the analysis of QTL underlying mean 100-seed weight at six different developmental stages. QTL x Environment interactions (QE) were analyzed by a mixed genetic mode based on 3 years' data. At an experiment-wise threshold of a=0.05 and by single-point analysis 94 QTL unaffected by QE underlay the mean seed weight at different developmental stages. Sixty-eight QTL affected by QE that also underlay mean seed weight were identified. From the 162 QTL 42 could be located on 12 linkage groups by composite interval mapping (LOD>2.0). The numbers, locations and types of the QTL and the genetic effects were different at each developmental stage. On linkage group C2 the distantly linked QTL swC2-1, swC2-2 and swC2-3 each affected mean seed weight throughout the different developmental stages. The DNA markers linked to the QTL possessed potential for use in marker-assisted selection for soybean seed size. The identification of QTL with genetic main effects and QE interaction effects suggested that such interactions might significantly alter seed weight during seed development.     

Antioxidant enzymes and isoflavonoids in chilled soybean (Glycine max (L.) Merr.) seedlings

Changes of activity antioxidant enzymes and of levels of isoflavonoids were studied in the roots and hypocotyls of the etiolated soybean (Glycine max (L.) Merr. var. Essor) seedlings, submitted to cold. Prolonged exposure to 1 degrees C inhibited hypocotyl and root elongation and limited their growth after seedlings were transferred to 25 degrees C. Roots were more sensitive to chilling than hypocotyls. At 1 degrees C a gradual increase in MDA concentration in roots but not in hypocotyls was observed. An increase in catalase (CAT, EC 1.11.1.6) and superoxide dismutase (SOD, EC 1.15.1.1) activity in hypocotyls was observed both at 1 degrees C and after transfer of plants to 25 degrees C. In roots, CAT activity increased after 4 days of chilling, while SOD activity only after rewarming. L-Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) activity decreased in roots of chilled seedlings, but did not change in hypocotyls until activity increased after transfer to 25 degrees C. The content of genistein and daidzein increased after 24 h of treatment by low temperature and then decreased with prolonged chilling in hypocotyls and remained high in roots. However, it should be noted that genistin level (genistein glucoside) in chilled hypocotyls is 10 times higher than in roots, despite falling tendency. The role of antioxidant enzymes and isoflavonoids in preventing chilling injury in hypocotyls and roots of soybean seedlings is discussed.