等渗盐胁迫下Na^＋和Cl^-对大豆幼苗光合作用的离子效应

研究和比较了等渗(-0.53MPa)的PEG-6000、NaCl、钠盐(无Cl-)和氯化物(无Na )溶液处理6d对栽培大豆品种‘Lee68’(耐盐性较强)和‘N23674’(耐盐性较弱)幼苗光合作用的离子效应。结果表明:PEG-6000处理使两品种叶片叶绿素含量和Rubisco活性较对照低,但降幅不如同样渗透压的NaCl、钠盐(无Cl-)和氯化物(无Na )溶液明显。PSII最大光化学效率(Fv/Fm)、电子传递速率(ETR)和PSII光化学的有效量子产额(Fv'/Fm')在PEG-6000处理2d和6d时显著下降,但在3种等渗盐处理下,多显著下降。两品种叶片气孔导度(Gs)和净光合速率(Pn)在4种胁迫处理下均显著下降,其中在3种盐处理下更明显,但胞间CO2浓度(Ci)仅在PEG-6000处理时下降,在盐处理下反而升高。两品种叶片叶绿素含量、Rubisco活性、Fv/Fm、ETR、Fv'/Fm'、Pn、Gs等在氯化物(无Na )溶液处理的下降幅度和叶绿体中Cl-含量及其与Na 总量的增加幅度均大于钠盐(无Cl-)处理的,在耐盐性弱的‘N23674’品种中更明显。可见,在NaCl胁迫对栽培大豆幼苗光合作用的毒害效应中,渗透胁迫较轻,离子毒害较重,其中Cl-的毒害大于Na 的。     

燕麦幼苗对盐胁迫的响应及过氧化氢对响应的调节

为了探讨‘定莜6号’燕麦对盐胁迫的生理生化响应及H2O2的调节作用,采用水培方法,研究外源H2O2对盐胁迫下燕麦活性氧代谢、渗透溶质积累和Na+、K+平衡的影响。结果表明:小于100 mmol·L-1Na Cl未对‘定莜6号’幼苗的生长造成明显影响,150 mmol·L-1及以上浓度Na Cl使幼苗干重和叶片K+/Na+显著降低,O2-·产生速率、H2O2、丙二醛(MDA)、可溶性蛋白质和脯氨酸含量及过氧化氢酶(CAT)、质膜H+-ATP酶活性明显提高,但抗坏血酸(ASA)和谷胱甘肽(GSH)含量变化不大;外施5μmol·L-1H2O2可显著缓解150 mmol·L-1Na Cl胁迫对燕麦幼苗生长的抑制作用,使燕麦叶片超氧化物歧化酶(SOD)、抗坏血酸过氧化物酶(APX)和CAT活性及H2O2、GSH含量明显提高,O2-·产生速率和MDA含量显著降低;5μmol·L-1H2O2还提高了150 mmol·L-1Na Cl胁迫下燕麦叶片可溶性蛋白质、可溶性糖、有机酸和脯氨酸含量及质膜H+-ATP酶活性和K+/Na+,降低了游离氨基酸含量;表明外源H2O2可调控燕麦幼苗活性氧代谢和渗透溶质积累,维持K+、Na+平衡,从而增强耐盐性。     

NaC1胁迫对4种豆科树种幼苗生长和K+、Na+含量的影响

以合欢、刺槐、国槐和皂英4种豆科树种盆栽实生幼苗为试验材料,研究了NaC1胁迫下4个树种幼苗的生长、耐盐临界浓度和Na+、K+含量的变化,并对其耐盐性进行了比较.结果表明:NaC1胁迫抑制了4个树种幼苗的生长,苗木的干物质积累量减小、根冠比增大,尤其对合欢和皂荚的影响较大;以相对干质量降至对照组50%时的NaC1浓度作为生长临界NaC1浓度(C50)指标,4个树种的耐盐强弱顺序为:刺槐(5.0‰)＞国槐(4.5‰)＞皂荚(3.9‰)＞合欢(3.0‰);随NaC1浓度的增加,各树种幼苗根、茎、叶中Na+含量逐渐增加,K+含量先增加后减小(合欢根除外),而K+/Na+差异较大.相同浓度NaC1胁迫下,幼苗器官的Na+分布为根＞茎＞叶,K+因树种和NaC1浓度不同而各异,以叶片中较多,K+/Na+为叶＞茎＞根.NaC1胁迫下,刺槐的K+含量和K+/Na+较高,地上部分Na+含量较低,幼苗干物质量大,耐盐性较强;而合欢的K+/Na+较小,高浓度NaC1胁迫下地上部分的Na+含量较高,幼苗干物质量小,耐盐性较差.苗木地上部分对K+的积累和根部对Na+的滞留是影响豆科树种耐盐性能的主要因素.     

NaCl胁迫对4种豆科树种幼苗生长和K~+、Na~+含量的影响

以合欢、刺槐、国槐和皂荚4种豆科树种盆栽实生幼苗为试验材料,研究了NaCl胁迫下4个树种幼苗的生长、耐盐临界浓度和Na+、K+含量的变化,并对其耐盐性进行了比较.结果表明:NaCl胁迫抑制了4个树种幼苗的生长,苗木的干物质积累量减小、根冠比增大,尤其对合欢和皂荚的影响较大;以相对干质量降至对照组50%时的NaCl浓度作为生长临界NaCl浓度(C50)指标,4个树种的耐盐强弱顺序为:刺槐(5.0‰)>国槐(4.5‰)>皂荚(3.9‰)>合欢(3.0‰);随NaCl浓度的增加,各树种幼苗根、茎、叶中Na+含量逐渐增加,K+含量先增加后减小(合欢根除外),而K+/Na+差异较大.相同浓度NaCl胁迫下,幼苗器官的Na+分布为根>茎>叶,K+因树种和NaCl浓度不同而各异,以叶片中较多,K+/Na+为叶>茎>根.NaCl胁迫下,刺槐的K+含量和K+/Na+较高,地上部分Na+含量较低,幼苗干物质量大,耐盐性较强;而合欢的K+/Na+较小,高浓度NaCl胁迫下地上部分的Na+含量较高,幼苗干物质量小,耐盐性较差.苗木地上部分对K+的积累和根部对Na+的滞留是影响豆科树种耐盐性能的主要因素.     

一氧化氮对裸燕麦耐盐性的增强效应

采用营养液培养方法,以‘定莜6号’裸燕麦为材料,研究外源一氧化氮供体硝普钠(SNP)对100 mmol·L-1Na Cl胁迫下裸燕麦幼苗生长、活性氧代谢和渗透溶质积累的影响。结果表明:5μmol·L-1SNP能明显缓解Na Cl胁迫对幼苗生长的抑制作用,显著提高Na Cl胁迫下裸燕麦叶片超氧化物歧化酶、过氧化物酶和抗坏血酸过氧化物酶活性及谷胱甘肽和抗坏血酸含量,降低丙二醛、过氧化氢、超氧阴离子和游离氨基酸含量及过氧化氢酶活性,提高可溶性糖、可溶性蛋白质和脯氨酸含量及K+/Na+比。分析表明,外源一氧化氮通过提高抗氧化能力和渗透溶质积累以及维持K+、Na+平衡,缓解盐胁迫诱导的氧化伤害和生长抑制,从而提高裸燕麦的耐盐性。     

盐胁迫对药用蒲公英生长特性及有效成分积累与离子吸收分配效应的影响

以药用蒲公英(Taraxacum officinale)为试材,研究不同浓度盐胁迫对其生长特性、有效成分积累和离子吸收分配的影响。结果表明,低盐胁迫(0.1% NaCl)对药用蒲公英生长和菊苣酸含量无显著影响,叶中Na+含量与对照无显著差异,K+含量及K+/Na+显著升高;高盐胁迫(≥0.2% NaCl)下其生长受到显著抑制,菊苣酸含量显著降低,类囊体膜结构随着盐胁迫加剧趋于紊乱,光合能力减弱,叶片Na+含量显著上升,而K+、Ca2+和Mg2+含量下降,K+/Na+、Ca2+/Na+和Mg2+/Na+显著降低。离子运输选择性系数(SCa,Na、SMg,Na、SK,Na)随着盐胁迫加剧呈先升后降趋势。相关性分析表明,盐胁迫下蒲公英叶片Na+含量与叶片生理指标呈极显著负相关。因此,叶片Na+富集是药用蒲公英遭受盐害导致生长受抑制的主要原因之一。     

盐生野大豆的异黄酮积累及其生态学意义

以自然生长在盐碱地上的野大豆(Glycine soja)和不耐盐的栽培大豆(G. max)为材料,测定了它们在不同盐度条件下叶片、根部和种子的异黄酮含量,并测定了它们叶片的L-苯丙氨酸含量和苯丙氨酸裂解酶(PAL)活性,还测定了它们根部的结瘤量和固氮酶活性。通过两者比较,分析了它们的大豆异黄酮代谢和盐渍环境的关系。结果表明:盐渍处理不抑制盐生野大豆PAL酶的活性,其大豆异黄酮大量积累;相反,盐渍处理明显抑制栽培大豆PAL酶活性,其大豆异黄酮含量减少,而大豆异黄酮合成前体L-苯丙氨酸积累。结果还显示:在盐渍条件下,盐生野大豆根部异黄酮积累的同时,其根瘤结瘤量较多,且固氮酶活性也较高;而栽培大豆随着其根部异黄酮的减少,其根瘤结瘤量大大减少,且固氮活性大大下降。野大豆和栽培大豆的这些差别说明:盐生野大豆积累大豆异黄酮有其生态学意义,这很可能是野大豆通过异黄酮次生代谢途径适应盐渍环境的一种重要机制。     

盐胁迫下水稻苗期Na+和K+吸收与分配规律的初步研究

选择苗期耐盐性较强的水稻(Oryza sativa)品种(株系)'AB52'、'02402'和'02435'及敏感品种'日本晴',在网室周转箱内,设置5 000和8 000 mg·L-1NaCl两种盐处理,以清水为对照,研究盐胁迫下苗期水稻植株不同部位Na+和K+的吸收和分配与品种耐盐性的关系.结果表明,盐胁迫下,株高、绿叶干重和绿叶面积下降,绿叶中的水分含量降低,但茎鞘中的水分含量有所上升.5 000 mg·L-1NaCl胁迫处理10 d,耐盐品种所受的生长影响和叶片伤害程度低于敏感品种,但8 000 mg·L-1NaCl胁迫处理下品种间差异变小.盐胁迫下,水稻植株吸收Na+和置换出K+,但不同器官部位中Na+和K+的区域化分布特征明显,各部位的Na+含量由低到高依次为绿叶、根、茎鞘和枯叶.下部老叶能优先积累较多Na+而枯黄;绿叶吸收Na+相对较少,维持较低的Na+水平,同时保持较高且稳定的K+含量;植株茎鞘通过选择性吸收大量Na+和置换出一部分K+到叶片中,保持绿叶较稳定的K+含量和相对较低的Na+含量,维持较高的K+/Na+比,从而使植株少受盐害.敏感品种'日本晴'在盐胁迫下绿叶中的Na+含量相对较高,且5 000 mg·L-1NaCl胁迫下绿叶Na+含量已接近高值,与在8 000 mg·L-1NaCl胁迫下差异不大,而耐盐品种绿叶吸收较少的Na+.另一方面,耐盐品种茎鞘的含K+相对较高,在盐胁迫下能吸收容纳较多的Na+,而绿叶中K+/Na+比较高.可以认为,绿叶的K+/Na+比可作为一个衡量耐盐性的相对指标.     

盐胁迫对一年生盐生野大豆幼苗活性氧代谢的影响

以国际上通用的栽培大豆品种Lee68(耐盐性较强)和非盐生野大豆N23232种群(耐盐性较弱)为参照材料,研究了在低盐(150mmol·L-1NaCl)和高盐(300mmol·L-1NaCl)胁迫下,盐生野大豆BB52种群幼苗体内的O-·2水平和MDA含量,活性氧清除酶系统中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)等酶活性,以及抗坏血酸(AsA)、类胡萝卜素(Car)、谷胱甘肽(GSH)等活性氧清除物质含量的变化。结果表明,在150和300mmol·L-1NaCl胁迫2d时,盐生野大豆BB52种群幼苗根和叶中的O-·2产生速率下降,SOD活性上升,其中叶片表现明显,且叶中APX活性也有增加。在不同浓度NaCl胁迫下,根中MDA含量均未见明显上升,叶片中略有增加。叶中AsA、Car和GSH含量均上升,且升幅相对较大。     

NaCl胁迫下棉花体内 Na~+ 、K~+分布与耐盐性

采用盐化土壤方法 ,选择苗期耐盐性较强的陆地棉品种枝棉 3号和中棉所 1 9及耐盐性较弱的品种泗棉 2号和苏棉 1 2号 ,研究了盐胁迫下棉苗体内 Na+、K+的运输和分配与耐盐性的关系。结果表明 ,耐盐品种根系具有一定的截留 Na+作用。棉花地上部盐分器官水平上的区域化分布特征明显 :2 0 0 mmol/L Na Cl胁迫下 ,枝棉 3号叶片中的 Na+含量显著低于泗棉 2号 ,茎及叶柄中的 Na+含量显著高于泗棉 2号 ;棉株地上部茎、叶柄、叶片中的 Na+含量分别由下而上逐渐减小 ,相同节位的茎、叶柄中的 Na+含量大于叶片 ,枝棉 3号更显著。1 0 0 mmol/L和 1 50 mmol/L Na Cl胁迫下 ,枝棉 3号和中棉所 1 9K+/Na+显著高于泗棉 2号和苏棉 1 2号。Na+在茎和叶柄中滞留和积累 ,根中的 K+向地上部选择性运输 ,以维持叶片中较高的 K+/Na+,是棉花耐盐性的一个重要特点     

Non-target impacts of soybean rust fungicides on the fungal entomopathogens of soybean aphid

Soybean aphid, Aphis glycines, has caused serious economic damage to soybean across the North Central US since its introduction to North America in 2000. The management of another invasive soybean pest, Asian soybean rust, Phakopsora pachyrhizi, using foliar fungicide applications has the potential to impact soybean aphid populations by suppressing beneficial fungal entomopathogens. In 2005 and 2006, we applied recommended soybean rust fungicide treatments, consisting of strobilurin and triazole fungicides, to small soybean plots in two locations to assess if such applications might suppress aphid fungal epizootics. In Lamberton, MN, in 2005, during the epizootic, fungicide-treated plots averaged 2.0 ± 0.7% (mean ± SE) disease prevalence while untreated plots averaged 14.2 ± 5.6%. In 2007, we applied strobilurin and strobilurin-triazole mix fungicides to single-plant microplots either before or after release of Pandora neoaphidis, the most commonly observed aphid pathogen in 2005 and 2006. Treatments that contained a mixture of two active ingredients significantly lowered peak and cumulative aphid disease prevalence in both early and late reproductive stage soybeans indicating that fungicide mixtures used to manage soybean rust can negatively impact an aphid-specific fungal pathogen. However, no consistent soybean aphid population response was observed in these studies of low levels of aphid fungal infection.     

Pseudomonas asturiensis sp. nov., isolated from soybean and weeds

Five strains of gram negative bacteria, isolated from soybean (LPPA 221^T, 222 and 223) and weeds (LPPA 816 and 1442), were analyzed by a polyphasic approach. The isolates showed variation in their phenotypic traits and were placed in the Pseudomonas fluorescens lineage, based on 16S rRNA gene sequence phylogeny, as a single but well separated cluster. MLSA analysis based on gyrB and rpoD sequences clustered the strains in a single branch in the Pseudomonas syringae group, and revealed P. viridiflava as closest relative. DNA–DNA hybridizations showed medium levels of DNA–DNA relatedness with the type strain of P. viridiflava (50%) and lower levels (<32%) with other type strains of the P. syringae group, supporting classification within a novel species of the genus Pseudomonas. The strains can be distinguished from species of the P. syringae group by the fatty acid C_17:0 cyclo that is present in a low amount (2.5%) and from P. viridiflava by their inability to assimilate d-tartrate and d-sorbitol, and by the formation of red colonies on TTC medium. For this new species, the name Pseudomonas asturiensis sp. nov. is proposed. The type strain is LPPA 221^T (=LMG 26898^T = CECT 8095^T).     

Molecular species of glycinin in some soybean cultivars

A(4) polypeptide-containing (Shirotsurunoko and York) and A(4) polypeptide-lacking (Raiden and Suzuyutaka) soybean cultivars were used to investigate the heterogeneity of glycinin molecular species. Purification of glycinin by DEAE-Toyopearl column chromatography afforded molecular species eluting before the glycinin fraction. Analysis of this fraction by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and sucrose density gradient centrifugation indicated that this protein consisted of A(1) and A(2) polypeptides. The A(4)-containing soybean cultivars contained less of this protein than the A(4)-lacking soybean cultivars, as exhibited by the size of the early peak appearing during column chromatography. Alkaline PAGE and N-terminal amino acid sequence analysis confirmed that the A(1)- and A(2)-rich molecular species in the A(4) polypeptide-lacking cultivars consisted of the A(1a) and A(2) polypeptides. Estimation of the molecular mass by gel permeation chromatography and multi-angle laser light scattering (GPC-MALLS) indicated that the A(1a)- and A(2)-rich molecular species were similar to a monomer of glycinin.     

Proteomic analysis of soybean nodule cytosol

An isolation procedure for soybean (Glycine max L. cv Williams 82) nodule cytosol proteins was developed which greatly improved protein resolution by two-dimensional polyacrylamide gel electrophoresis. The most abundant proteins were selected and analyzed by mass spectrometry. The identified proteins were categorized by function (% of total proteins analyzed): carbon metabolism (28%), nitrogen metabolism (12%), reactive oxygen metabolism (12%) and vesicular trafficking (11%). The first three categories were expected based on the known physiological functions of the symbiotic nitrogen fixation process. The number of proteins involved in vesicular trafficking suggests a very active exchange of macromolecules and membrane components. Among the 69 identified proteins were the enzymes of the three carbon portion of glycolysis, which were further characterized to support their roles in the sucrose synthase pathway to provide malate for the bacteroids. Proteomic analysis provides a functional tool by which to understand and further investigate nodule function.     

Analysis of polyamine metabolism in soybean seedlings using 15N-labelled putrescine

The translocation and metabolism of polyamines during soybean germination were studied using 15N-labelled putrescine as a precursor. Both 15N-labelled and unlabelled polyamines were simultaneously detected using a novel application of ionspray ionization-mass spectrometry. 15N-putrescine was rapidly transported to the shoots and roots, where it was converted to spermidine and spermine. The main 15N-polyamine that accumulated in the root was 15N-spermine. It was found that there were differences in the way endogenous putrescine and exogenous 15N-putrescine were metabolized in soybean seedlings.     

Identification of the soybean HyPRP family and specific gene response to Asian soybean rust disease

Soybean [Glycine max (L.) Merril], one of the most important crop species in the world, is very susceptible to abiotic and biotic stress. Soybean plants have developed a variety of molecular mechanisms that help them survive stressful conditions. Hybrid proline-rich proteins (HyPRPs) constitute a family of cell-wall proteins with a variable N-terminal domain and conserved C-terminal domain that is phylogenetically related to non-specific lipid transfer proteins. Members of the HyPRP family are involved in basic cellular processes and their expression and activity are modulated by environmental factors. In this study, microarray analysis and real time RT-qPCR were used to identify putative HyPRP genes in the soybean genome and to assess their expression in different plant tissues. Some of the genes were also analyzed by time-course real time RT-qPCR in response to infection by Phakopsora pachyrhizi, the causal agent of Asian soybean rust disease. Our findings indicate that the time of induction of a defense pathway is crucial in triggering the soybean resistance response to P. pachyrhizi. This is the first study to identify the soybean HyPRP group B family and to analyze disease-responsive GmHyPRP during infection by P. pachyrhizi.     

Effect of chaotropic agents on reversible unfolding of a soybean (Glycine max) seed acid phosphatase

In this work we examined the effect of urea and guanidinium chloride on the structural stability of a single isoform of soybean seed acid phosphatase, based on the intensity of tryptophan fluorescence as a function of denaturant concentration. The free energy of unfolding, DeltaGu, was calculated at 25 degrees C as a function of the concentrations of both chaotropic agents; the conformational stability, DeltaG (H2O), was determined to be 2.48 kcal mol(-1). Center of mass, determined from analysis of fluorescence data, was used as a parameter to assess conformational changes. Our results indicate that complete enzyme inactivation occurred before full enzyme unfolding in both cases, and suggest that there are differences between the conformational flexibility of the active-site and that of the macromolecule as a whole.     

Taxonomical re-evaluation of Phoma-like soybean pathogenic fungi

Coelomycetous fungi classified in Ascochyta, Phoma, and Phyllosticta have been recorded from spots on leaves and pods of soybeans. Based on the Genealogical Concordance Phylogenetic Species Concept, the authors suggest the re-evaluation of the taxonomic status of Phoma sojicola (syn. = Ascochyta sojicola) and Phyllosticta sojicola. In spite of the former delimitation of Phoma sojicola based on small differences in morphological features, it has proved to be identical to Phoma pinodella. Similarly, it was also confirmed that Phyllosticta sojicola was identical to Phoma exigua var. exigua. The authors supply tools for identification of Phoma-like fungi by combined conventional and molecular methods. Protein-encoding genes (tef1 and β-tubulin) were successfully applied within the Phoma genus to infer phylogenetic relationships.     

Aryl hydroxylation of isopropyl-3-chlorocarbanilate by soybean plants

Isopropyl-3-chlorocarbanilate-phenyl UL-¹⁴C (CIPC-¹⁴C) is absorbed, translocated and metabolized by soybean plants. Water-soluble metabolites in root and shoot were purified and the root major metabolite characterized. The acetylated aglucones from the β-glucosidase hydrolysis and the esters from the direct acetylation of CIPC-¹⁴C polar metabolites were purified by GLC and analysed by mass spectrometry. The data showed that the phenyl riong of CIPC-¹⁴C was hydroxylated by both root and shoot tissues. Isopropyl-5-chloro-2-hydroxycarbanilate (hydroxy-CIPC) was the predominant aglucone liberated by β-glucosidase from polar metabolites in root and shoot. The o-glucoside of hydroxy-CIPC was shown to be present, by direct acetylation and characterization. In shoot tissue the major metabolites were dechlorinated hydroxy-CIPC and were not hydrolysed by β-glucosidase. These data show that soybean root or shoot tissues hydroxylate the phenyl ring of CIPC-¹⁴C but do not alter either the isopropyl alcohol moiety or the earbamate bond.