低铁胁迫对玉米苗期根系生长和铁素吸收利用的影响

为了揭示不同耐低铁玉米品种苗期根系生长和铁素吸收利用的差异,为玉米耐低铁能力的遗传改良提供依据,以耐低铁玉米品种‘正红2号’和不耐低铁玉米品种‘川单418’为材料,采用重度[10μmol(Fe~(3+))·L~(-1)]、中度[30μmol(Fe~(3+))·L~(-1)]和轻度[50μmol(Fe~(3+))·L~(-1)]3种低铁胁迫及对照[100μmol(Fe~(3+))·L~(-1)]的铁营养液处理3叶1心玉米幼苗,分析低铁胁迫对不同耐低铁玉米品种苗期根系生长和铁素吸收利用的影响。结果表明,随着营养液铁浓度降低,两个玉米品种幼苗的根长、根体积、根系活力、干物质、铁含量、铁积累量、相对吸铁能力均显著降低,但根系麦根酸分泌量增多,铁素向地上部转移分配能力增强,铁素的生理效率提高,这是玉米适应低铁胁迫的重要生理机制之一。玉米幼苗的铁素积累量与根长、根体积、根干重、根系活力等根系性状均呈极显著或显著正相关。耐低铁玉米品种在中度和重度低铁胁迫下根长、根体积、根干重、根系活力均较不耐低铁玉米品种高,是其铁素吸收积累量高的重要原因。根系麦根酸分泌量与铁素茎叶分配率呈正相关,铁素茎叶分配率与铁素生理效率呈极显著正相关,增加根系麦根酸的分泌量可在一定程度上提高玉米铁素的茎叶分配率,从而提高铁素生理效率;耐低铁玉米品种在中度和重度低铁胁迫下麦根酸分泌量增幅高于不耐低铁玉米品种,是其铁素生理效率高的主要原因。     

苯丙烯酸胁迫对黄瓜幼苗生长和抗氧化代谢特性的影响

采用营养钵土培的方法,研究了外源苯丙烯酸对津美1号、绿衣天使、C90和翠龙等不同黄瓜品种幼苗生长和叶片抗氧化酶活性及活性氧代谢的影响。结果表明,外源苯丙烯酸胁迫对黄瓜幼苗生长具有很强的抑制作用,并随着胁迫浓度的增大受抑制的程度加强。在苯丙烯酸胁迫下,黄瓜叶片的超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）和抗坏血酸过氧化物酶（APX）活性表现为低浓度升高,高浓度下降的趋势;超氧阴离子（O2-.）产生速率、过氧化氢（H2O2）含量、膜脂过氧化产物丙二醛（MDA）含量及渗透调节物质脯氨酸（Pro）含量、可溶性糖含量随着苯丙烯酸浓度的升高而显著增加。不同黄瓜品种对苯丙烯酸胁迫的响应存在着品种间差异。     

硝普钠对黄瓜幼苗缺镁和硝酸盐胁迫的缓解效应

【目的】设施栽培中土壤次生盐渍化严重,导致植株出现缺镁失绿症状。通过研究硝酸盐胁迫下外源NO供体(硝普钠,SNP)对缺镁黄瓜幼苗生长的影响,探究硝酸盐胁迫下外源NO对缺镁黄瓜幼苗的胁迫缓解效应,为解决设施生产中黄瓜幼苗的缺镁失绿症状提供理论指导。【方法】采用水培的方式培养黄瓜幼苗,幼苗长至三叶一心时进行处理。营养液采用山崎配方,镁离子浓度设两个水平为2 mmol/L(正常浓度)和1 mmol/L(缺镁胁迫);硝酸盐浓度设两个水平为14 mmol/L(正常浓度)和140 mmol/L(硝酸盐胁迫);硝酸盐和镁正常浓度为对照。用0.1mmol/L SNP分别缓解缺镁胁迫、硝酸盐胁迫以及缺镁和硝酸盐双重胁迫,用0.1 mmol/L SF(铁氰化钠)处理,观察SNP反应产物的影响;NO-3由Ca(NO3)2·4H2O和KNO3提供,各占1/2,p H由H2SO4调节,保持在5.5 6.5。【结果】1)缺镁胁迫下,黄瓜幼苗株高和叶面积增加值、干物质增长量较正常处理的黄瓜幼苗显著降低;电解质渗漏率、丙二醛含量和可溶性蛋白含量较正常处理的黄瓜幼苗显著升高。缺镁处理的黄瓜幼苗根茎叶中镁离子含量、叶片光合色素含量、光合特性指标、叶绿素荧光及抗氧化酶活性与正常处理的黄瓜幼苗相比明显降低。2)硝酸盐胁迫下,黄瓜幼苗株高和叶面积增加值、干物质增长量、黄瓜幼苗根茎叶中镁离子含量、叶片光合色素含量、光合特性指标、叶绿素荧光及抗氧化酶活性较正常处理的黄瓜幼苗显著降低;电解质渗漏率、丙二醛含量和可溶性蛋白含量较正常处理的黄瓜幼苗显著升高。3)缺镁和硝酸盐双重胁迫下,黄瓜幼苗的相关生长指标和抗逆指标较正常处理的黄瓜幼苗降低或增大更为显著。4)缺镁胁迫,硝酸盐胁迫以及缺镁和硝酸盐双重胁迫下,外施0.1 mmol/L SNP处理的黄瓜幼苗,株高和叶面积增加值、干物质增长量、幼苗根茎叶中镁离子含量、叶片光合色素含量、光合特性指标、叶绿素荧光、可溶性蛋白含量及抗氧化酶活性较未添加SNP处理的黄瓜幼苗显著提高,电解质渗漏率和丙二醛含量则明显降低。外施0.1 mmol/L SF则没有表现出明显的作用。【结论】硝酸盐胁迫下缺镁黄瓜幼苗生长受到明显抑制,出现失绿症状,通过添加0.1 mmol/L SNP,黄瓜幼苗的生长抑制得到明显缓解,说明在硝酸盐胁迫下外源NO对缺镁黄瓜幼苗的胁迫有显著缓解作用,增强黄瓜幼苗的耐盐性和对镁的吸收能力。     

高浓度营养液对黄瓜和番茄下胚轴徒长的抑制作用

【目的】黄瓜和番茄是我国主栽蔬菜种类,育苗过程中容易遭遇高温、高湿、弱光等不良环境,导致幼苗下胚轴徒长,严重影响后期丰产性。在控制幼苗徒长的实际操作中,利用植物生长调节剂、机械刺激、补光、温度调节、水分控制等调控方法容易受到多种因素影响而效果不佳。本文研究高浓度营养液浓度对黄瓜和番茄幼苗下胚轴徒长的抑制作用,以期提出一种有效制御幼苗徒长的方法。【方法】试验以Hoagland标准营养液配方为基础浓度（设为1.0C）,通过成比例增减大量元素组分用量而不改变微量元素组分用量,组成0.5C、2.5C、5.0C、7.5C、10.0C和12.5C 6个浓度。以0.5C为对照,在日光温室条件下分别对黄瓜和番茄幼苗进行灌施处理。待幼苗生长至子叶平展期时,测定黄瓜和番茄幼苗形态及生长指标,研究营养液浓度对幼苗下胚轴细胞长度、生长速率及内源激素含量的影响;随后对幼苗进行0.5C低浓度营养液的恢复生长处理,待幼苗生长至第一真叶平展期时,再次进行幼苗形态及生长指标的测定,从而确定高浓度营养液对黄瓜和番茄下胚轴徒长的抑制效果及机制,并确定最适合制御幼苗徒长的营养液浓度。【结果】6种浓度营养液对黄瓜和番茄幼苗下胚轴徒长的影响结果为: 1）高浓度营养液显著提高了基质EC值（r=0.99）,黄瓜和番茄幼苗在子叶平展期其下胚轴生长受到明显抑制,下胚轴长度、绝对生长速率和含水量均呈下降趋势,下胚轴皮层薄壁细胞轴/径向生长也受到影响。与对照相比,10.0C营养液处理下黄瓜和番茄幼苗下胚轴皮层薄壁细胞缩短为49%和48%。2）随着营养液浓度的增加,黄瓜和番茄下胚轴内源激素含量发生显著变化,其中赤霉酸（GA3）、玉米素核苷（ZR）含量呈降低趋势,而脱落酸（ABA）含量呈升高趋势,且ABA含量与下胚轴长度呈极显著负相关关系（P＜0.01）。生长素（IAA）含量在不同浓度营养液处理下呈现区位性变化特征。3）在幼苗子叶平展期重新灌施0.5C浓度的营养液,对5.0C和7.5C处理下生长的幼苗均有恢复生长的作用,且幼苗株高/茎粗比明显优于对照和其它处理,该浓度可望有效避免幼苗徒长。【结论】高营浓度养液通过改变幼苗内源激素的动态平衡,特别是促进ABA的合成和积累,从而抑制幼苗下胚轴皮层细胞伸长生长,有效制御幼苗徒长。灌施高浓度营养液对黄瓜、番茄幼苗下胚轴伸长的抑制作用可以通过降低营养液浓度而得到解除,这种防治幼苗徒长的操作方法简便易行,具有潜在的应用前景,但在不同品种蔬菜幼苗生长过程中,最优化的营养液施用浓度及其影响机制尚需进一步研究分析。     

梨树内生细菌LP-5的鉴定及其促生作用研究

从梨树木质部分离到1株具有强烈抑菌活性的内生细菌LP-5,通过形态观察、生理生化测定、16SrDNA序列分析及特异基因片段扩增,将其鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。盆栽试验结果表明,该菌株可明显促进黄瓜幼苗的生长,其20倍稀释发酵液处理黄瓜种子,可使其胚根长度增长36.32%。用同稀释倍数的发酵液处理黄瓜幼苗,可使幼苗株高、茎粗、干重分别提高9.06%、14.67%和24.14%。高效液相色谱分析表明:该菌株具有促生作用可能是因为产生植物激素IAA。     

外源谷胱甘肽对自毒作用下嫁接黄瓜及砧穗幼苗叶片抗氧化系统的影响

为探索外源还原型谷胱甘肽(GSH)对自毒作用下嫁接黄瓜的保护作用及机理,本研究以200g·L-1连作嫁接黄瓜根际土壤浸提液灌根模拟自毒胁迫(CK2),分别以浓度为0、100、200 mg·L-1GSH对幼苗进行叶片喷施和灌根处理,在处理第4、第8、第12、第16天测定嫁接黄瓜、黄瓜、南瓜幼苗生物量、叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)和过氧化氢酶(CAT)活性以及GSH、抗坏血酸(As A)、丙二醛(MDA)含量。结果表明,自毒胁迫下,嫁接黄瓜、黄瓜和南瓜幼苗壮苗指数、叶片SOD、POD、APX、CAT活性降低,As A、GSH含量下降,MDA含量升高。表明连作嫁接黄瓜土壤浸提液能够对3种植物幼苗产生自毒作用。3种植物幼苗T1、T2、T3处理和嫁接黄瓜、南瓜幼苗T4处理的壮苗指数,叶片SOD、POD、APX、CAT活性和GSH、As A含量均显著增加,MDA含量均显著降低。黄瓜幼苗T4处理的壮苗指数,叶片SOD、POD、APX、CAT活性和GSH、As A含量呈下降趋势,MDA含量呈上升趋势。表明施加适宜浓度GSH可以提高抗氧化酶活性和抗氧化剂含量,减轻活性氧对膜脂造成的伤害,从而提高幼苗素质。综上,嫁接黄瓜和南瓜幼苗的T4处理,黄瓜幼苗的T2处理缓解自毒作用效果较为明显。本研究结果为防治嫁接黄瓜连作障碍提供了理论依据。     

旱作黄潮土中铁元素的有效性研究

通过对不同季节采集的不同质地类型土壤样品中有效铁、有机质和酸碱度的化验分析及对小麦、花生、黄瓜进行根外喷施 0 .5 %硫酸亚铁溶液对比研究 ,认为在旱作黄潮土中铁元素的有效性与土壤粘粒含量和有机质含量呈正相关 ,雨季土壤大于旱季土壤 ,而酸碱度的相关性不明显 ;花生、黄瓜对铁素营养的敏感性大于小麦。根外喷施铁肥应优先在质地较粗的旱季土壤和花生、黄瓜上应用     

产ACC脱氨酶植物根际促生菌的筛选及其对修复植物高羊茅生长的影响

以1-氨基环丙烷-1-羧酸(ACC)为唯一氮源,从石油污染的土壤中分离得到一株具有ACC脱氨酶活性的菌株D5A.该菌在以ACC为唯一氮源条件下,其ACC脱氨酶比活力为0.084 U/mg.另外,D5A还具有产生吲哚乙酸(IAA)、耐盐以及溶磷的特性.在液体培养条件下该菌株产IAA高达112 mg/L,在90 g/kg盐含量的培养基中仍能够正常生长且具有较强的溶解矿物磷能力.同时该菌对酸碱具有良好的适应性,在初始pH4～10的LB培养基中生长良好.种子发芽试验表明在3 g/kg和6 g/kg浓度NaC1的逆境条件下,该菌株能显著提高高羊茅种子的发芽率和芽长.最后,通过对其进行生理生化特性和16S rDNA序列分析,该菌株初步鉴定为克雷伯氏菌(Klebsiellasp.).     

不同温度下镁胁迫对黄瓜光合特性和活性氧清除系统的影响

利用人工气候箱,研究不同温度下镁胁迫对黄瓜幼苗光合特性和活性氧清除系统的影响。结果表明,不论温度高低,镁胁迫增加了叶绿素a/b,且缺镁比高镁增幅大。在两种温度下,镁均降低了叶绿素a、b和类胡萝卜素的含量;而在适温下,高镁使叶绿素a、b和类胡萝卜素含量增加,低温则降低。镁胁迫使黄瓜幼苗中下部叶片净光合率（Pn）、气孔导度（Gs）下降,低温、高镁Pn降低幅度最大;但对胞间CO2 浓度（Ci）、蒸腾速率（Tr）、水分利用率（WUE）和气孔限制值（Ls）的影响因温度和镁离子浓度不同而不同。缺镁胁迫,黄瓜幼苗光合作用的主要限制因素是非气孔因素,而高镁胁迫则主要限制因素是气孔因素。镁胁迫使黄瓜幼苗叶片MDA含量、O2-.产生速率、H2O2含量增加,缺镁增幅大于高镁胁迫;低温加剧了膜脂过氧化。低温下,POD活性下降,SOD活性升高,而CAT活性变化不大;适温下,植株主要通过提高POD活性来抵御镁胁迫。低温、缺镁对植株的伤害较大,与活性氧的增加有关。     

甘蔗内生菌B9的鉴定及其促生长机制和定殖能力的研究

B9是一株分离于甘蔗品种"云蔗99-91"根部的内生菌,笔者对其种属鉴定与促生长机制开展研究,以期应用于甘蔗生产。试验通过传统方法与16S rRNA等看家基因片段测序对菌株进行分类鉴定;采用固体透明圈、酸性钼蓝比色法、火焰分光光度法、GFP荧光标记等方法测定与研究B9菌株固氮、溶磷、解钾、产IAA、嗜铁素等生物学活性及在甘蔗苗内的定殖情况;最后通过回接法研究菌液处理对玉米种子萌发与甘蔗幼苗生长发育的影响。结果表明B9菌属于芽孢杆菌属枯草芽孢杆菌(Bacillus subtilis),具有固氮、溶磷、解钾、产生IAA和嗜铁素的能力。用B9菌液处理后能提高玉米种子的发芽势与促进根芽组织生长,同时处理组甘蔗苗株在农艺性状与生理生化水平上明显优于无菌清水对照,并且发现B9菌能在甘蔗根茎叶内稳定定殖。本文鉴定了B9菌的种类和功能,阐明了B9菌的促生长机制和定殖能力,为开发生物菌肥并应用于生产提供理论依据。     

多功能植物根际促生菌 DD3 的功能特性及对大蒜幼苗的促生效果

【目的】植物根际促生菌 (plant growth promoting rhizobacteria, PGPR) 能够促进植物生长、防治病害、增加作物产量,具有较好的环境兼容性、不易引起抗性、效果持久稳定等优势。筛选高效多功能菌,探索抗病促生机理,对开发防控大蒜土传病害的微生物技术,发展“高产、优质、高效、生态、安全”的大蒜产业具有十分重要的理论和现实意义。 【方法】以实验室前期从山东省金乡县连作大蒜根部分离的 70 株细菌为供试菌株,筛选对大蒜根腐病病原真菌拟枝孢镰刀菌 (Fusarium sporotrichioides) ACCC37402 和大蒜叶枯病病原真菌链格孢属菌株 (Alternaria sp.) D14011-1 具有较好拮抗效果的菌株;采用 16S rDNA 基因序列同源性分析,对其进行初步鉴定;通过测定菌株对抗生素的敏感性、水解蛋白能力、产铁载体能力、产 HCN 能力等,研究菌株的拮抗特性;通过测定菌株的固氮能力、溶磷能力、产 IAA 能力,产 ACC 脱氨酶能力等,研究菌株的促生特性;通过温室盆栽试验,进行多功能菌菌悬液及其无菌发酵液对大蒜幼苗生长的影响以及抗病效果研究。 【结果】菌株 DD3 对大蒜根腐病病原真菌 ACCC37402 和大蒜叶枯病病原真菌 D14011-1 具有较好拮抗效果,抑菌率均为 45%;经 16S rDNA 基因序列同源性分析,初步确定为芽孢杆菌属 (Bacillus) 菌株。其它功能特性研究结果显示:菌株 Bacillus sp. DD3 具有水解蛋白能力、产 HCN 能力、产铁载体能力、固氮能力、溶解有机磷和无机磷的能力;对氨苄青霉素钠、硫酸卡那霉素等 11 种抗生素无耐受性,不具有分泌吲哚乙酸 IAA 能力和产 ACC 脱氨酶能力。盆栽试验显示,接种 Bacillus sp. DD3 菌悬液对大蒜根腐病的防治效果较好,防控率达 80%。接种 Bacillus sp. DD3 及其无菌发酵液后能够促进大蒜幼苗的生长,其中接种菌悬液效果更佳。 【结论】菌株 Bacillus sp. DD3 兼具多种功能特性,对大蒜根腐病具有较好的拮抗效果,同时可以显著促进大蒜幼苗生长,有望进一步研究开发成为微生物肥料生产菌种。     

Effect of the expression of Escherichia coli fhuA gene in Rhizobium sp. IC3123 and ST1 in planta: Its role in increased nodule occupancy and function in pigeon pea

The inability to utilize a fungal siderophore as source of iron nutrition by most of the rhizobial cultures isolated from pigeon pea, could be considered a negative fitness factor since hydroxamate siderophores are found in significant amounts in natural soils. Thus these cultures were engineered to use ferrichrome a prototype of hydroxamate type siderophore. Pigeon pea Rhizobium spp. IC3123 and ST1 harboring Escherichia coli fhuA gene, responsible for uptake of Fe³⁺-ferrichrome, were obtained by transformation with pGR1, a broad host range plasmid carrying the fhuA gene under the control of the lac promoter of E. coli. Expression of fhuA in transformed rhizobial strains IC3123::pGR1 and ST1::pGR1 was confirmed by the ability of the plasmid-bearing strains to utilize iron bound to ferrichrome. Inoculation of pigeon pea plants with fhuA expressing rhizobial strains in pot experiments showed a significant increase in plant growth as well as nodule density as compared to those inoculated with the parent as well as the empty vector-bearing strain. Inoculation of pigeon pea seedlings with IC3123::pGR1 and ST1::pGR1 led to marked increase in shoot fresh weight, nodule number per plant, chlorophyll content of leaves and effective nodule symbiosis when compared with plants inoculated with the parent strains IC3123 and ST1. The positive effect of IC3123::pGR1 and ST1::pGR1 treatment on plant growth was more significantly observed when ferrichrome producing Ustilago maydis, known to secrete ferrichrome, was co-inoculated along with the transformed rhizobia. The presence of fhuA gene in rhizobial strains also led to an increased survival and root colonization.     

Plant growth-promoting bacterium Pseudomonas sp. strain GRP3 influences iron acquisition in mung bean (Vigna radiata L. Wilzeck)

Siderophores produced by Pseudomonas sp. may be used by the bacteria (homologously) or in effecting plant nutrition (heterologously). The problem of iron non-availability particularly in calcareous soils may be overcome by incorporation of siderophore producing strains of fluorescent psuedomonads (FLPs). Siderophore producing bacterium Pseudomonas strain GRP₃ was used in a pot experiment to assess the role of microbial siderophores in the iron nutrition of mung bean (Vigna radiataL. Wilzeck) using Fe-citrate, Fe-EDTA and Fe(OH)₃ in different concentrations with Hoagland's solution. After 45 days, the plants showed a reduction of chlorotic symptoms and enhanced chlorophyll level in GRP₃ bacterized plants. Bacterization with GRP₃ increased peroxidase activity and lowered catalase activity in roots. In 10 μM Fe-citrate alongwith GRP₃ treatment, chlorophyll a, chlorophyll b and total chlorophyll contents increased significantly by 34, 48 and 39%, respectively, compared to the control. Peroxidase activity in the same treatment was increased by 82% whereas catalase activity decreased by 33%. There was also a significant increase in total and physiologically available iron. A closely similar pattern was observed in chlorophyll content and peroxidase activity in Fe-EDTA and Fe(OH)₃ treated plants; catalase activity was an exception. The data suggests operation of heterologous siderophore uptake system in mung bean in presence of GRP₃. Such siderophore producing system has the potential of improving iron availability to plants and reduce fertilizer usage.     

Effect of the expression of Escherichia coli fhuA gene in Rhizobium sp. IC3123 and ST1 in planta: Its role in increased nodule occupancy and function in pigeon pea

The inability to utilize a fungal siderophore as source of iron nutrition by most of the rhizobial cultures isolated from pigeon pea, could be considered a negative fitness factor since hydroxamate siderophores are found in significant amounts in natural soils. Thus these cultures were engineered to use ferrichrome a prototype of hydroxamate type siderophore. Pigeon pea Rhizobium spp. IC3123 and ST1 harboring Escherichia coli fhuA gene, responsible for uptake of Fe³⁺-ferrichrome, were obtained by transformation with pGR1, a broad host range plasmid carrying the fhuA gene under the control of the lac promoter of E. coli. Expression of fhuA in transformed rhizobial strains IC3123::pGR1 and ST1::pGR1 was confirmed by the ability of the plasmid-bearing strains to utilize iron bound to ferrichrome. Inoculation of pigeon pea plants with fhuA expressing rhizobial strains in pot experiments showed a significant increase in plant growth as well as nodule density as compared to those inoculated with the parent as well as the empty vector-bearing strain. Inoculation of pigeon pea seedlings with IC3123::pGR1 and ST1::pGR1 led to marked increase in shoot fresh weight, nodule number per plant, chlorophyll content of leaves and effective nodule symbiosis when compared with plants inoculated with the parent strains IC3123 and ST1. The positive effect of IC3123::pGR1 and ST1::pGR1 treatment on plant growth was more significantly observed when ferrichrome producing Ustilago maydis, known to secrete ferrichrome, was co-inoculated along with the transformed rhizobia. The presence of fhuA gene in rhizobial strains also led to an increased survival and root colonization.     

玉米根际高效固氮菌Sphingomonas sp. GD542的分离鉴定及接种效果初步研究

固氮微生物菌种选育是固氮微生物肥料生产应用的基础。本研究从玉米根际土壤分离到1株高效固氮菌株GD542,菌体短杆状,0.4μm×1.0～1.5μm,革兰氏阴性;固氮酶活性为5.046nmol(C2H4)·h-1·mg-1(蛋白);利用碳源较广泛,抗逆性较强,既可在4℃低温生长,也可以在37℃下生长,耐盐性高达10%。16SrDNA序列分析结果表明,该菌株与鞘氨醇单孢菌Sphingomonas azotifigens的16SrDNA序列有高达96%的同源性,结合形态特征和生理生化特征等,将其初步鉴定为鞘氨醇单孢菌Sphingomonas sp.。接种小白菜的温室盆栽试验表明,菌株GD542具有很好的固氮效能,与无氮对照相比,接种GD542处理植株干重增加206%,含氮量增加230%,达到统计学显著差异水平,开发应用前景较好。     

适应玉米的溶磷细菌筛选及其对玉米生长的影响

从石灰性土壤中分离获得4株高效溶磷细菌X5、X6、Z4和Z8,研究其生物学特征,探索其单独及复合的溶磷促生潜能。研究发现菌株X5、X6、Z4和Z8均可以利用玉米根系分泌物作碳源生长。菌株X6和Z4均能产生吲哚乙酸(IAA)和铁载体,菌株Z8可产生IAA不产生铁载体,菌株X5可产生铁载体不产生IAA。盆栽试验结果表明,接种单一溶磷菌及4株菌复合处理均可促进玉米生长,但复合菌群的溶磷促生效果显著高于单一菌株。通过16S r RNA基因序列分析研究菌株的分类地位,初步鉴定X5、X6、Z4、Z8分别为荧光假单孢菌(Pseudomonas fluorescens)、草假单胞菌(Pseudomonas poae)、巨大芽孢杆菌(Bacillus megaterium)和枯草芽孢杆菌(Bacillus subtilis)。     

Importance of 2,4-DAPG in the biological control of brown patch by Pseudomonas fluorescens HP72 and newly identified genes involved in 2,4-DAPG biosynthesis

The Pseudomonas fluorescens strain HP72 used as biocontrol agent was isolated from the roots of creeping bentgrass on brown patch-suppressive soil. This strain can suppress brown patch disease caused by Rhizoctonia solani. The analysis of secondary metabolites from strain HP72 revealed that it produced known antifungal compounds, 2,4-diacetylphloroglucinol (2,4-DA-PG), HCN, and a fluorescent siderophore. In the present study, the Tn5inserted mutants of strain HP72, which did not show any antifungal activity, were selected. None of the mutants produced 2,4-DA-PG but they produced a fluorescent siderophore, while some strains produced HCN. Therefore, it is suggested that 2,4-DA-PG plays a major role in the biological control of brown patch disease caused by R. solani. In the genomic region where Tn5 was inserted, two open reading frames (ORFs A and B), which are not included in the 2,4-DAPG gene cluster of HP72, were detected. It was demonstrated that ORFs A and flare involved in the regulation of 2,4-DAPG biosynthesis.     

降解玉米秸秆真菌复合菌系的构建及其降解效果评价

  【目的】  秸秆的木质纤维素含量丰富、结构复杂，在自然界中降解较慢，增加秸秆降解菌剂中菌株的多样性有利于提升还田秸秆的降解效果。探究菌株多样性水平和组成影响复合菌系秸秆降解的效果及原因，为复合菌系在秸秆降解中的应用提供理论支撑。  【方法】  通过富集驯化培养，从玉米秸秆还田土壤中筛选具有秸秆降解能力的真菌，从中挑选5株高效秸秆降解真菌进行基因间隔区序列 (ITS) 测定和物种鉴定，明确其分类地位。通过全组合构建菌株多样性为1～5的复合菌系，分别检测复合菌系的秸秆相对降解率及其滤纸酶、纤维素内切酶和木聚糖酶活性，利用方差分析和相关性分析等方法研究菌株多样性和组成对复合菌系玉米秸秆降解效果及其纤维素酶活性的影响。  【结果】  共筛选获得了15株具有秸秆降解能力的真菌，其中5株真菌的秸秆降解效果好、纤维素水解能力强。经ITS序列鉴定和系统发育分析，发现5株降解真菌的遗传差异较大，Z7-6、F7-5、F4-3、L1-1和J2-5分别与草酸青霉 (Z7-6: Penicillium oxalicum)、烟曲霉 (F7-5: Aspergillus fumigatus)、哈茨木霉 (F4-3: Trichoderma harzianum)、白囊耙齿菌 (L1-1: Irpex lacteus) 和木贼镰刀菌 (J2-5: Fusarium equiseti) 的ITS序列相似度均超过99.95%。全组合复配结果表明，复合菌系的秸秆降解能力和纤维素酶活力均高于各单一菌株，且随着菌株多样性水平的增加而提高。滤纸酶、纤维素内切酶和木聚糖酶的活力越强，复合菌系对玉米秸秆的降解效果越好，而其秸秆相对降解率主要取决于滤纸酶和纤维素内切酶的活性。抽样效应分析发现，不同菌株对复合菌系的秸秆降解效果、滤纸酶和纤维素内切酶活性的影响不同。不含菌株F7-5的复合菌系降解效果显著优于含有该菌株的组合，以Z7-6 (P. oxalicum)、F4-3 (T. harzianum)、L1-1 (I. lacteus) 和J2-5 (F. equiseti) 组合F1的玉米秸秆降解效果最佳、酶活性最高。  【结论】  秸秆降解复合菌系的构建过程需要同时考虑多样性效应和抽样效应，增加降解菌的多样性有助于增强秸秆的降解效果。本研究筛选获得的复合菌系F1在玉米秸秆降解中具有潜在的应用前景。     

Efficacy of an artificial microbial siderophore-Fe(III) with high redox potential on correcting Fe chlorosis in rice

ABSTRACT

Microbial siderophore-chelated Fe(III) is suggested to be an important source of Fe for plants, although it is hardly reduced by plant roots. Here, we investigated the efficacy of the easily reducible artificial microbial siderophore tris[2-{(N-acetyl-N-hydroxy)glycylamino}ethyl]amine (TAGE)-Fe(III) as an alternative Fe source to correct Fe deficiency in rice plants, and compared it to that of the natural siderophore deferoxamine B (DFOB)-Fe(III). We also evaluated the absorption of Fe from TAGE-Fe(III) by the Strategy I-like system of gramineous plants using nicotianamine aminotransferase 1 (naat1) mutant rice, which does not synthesize phytosiderophores. Fe(III)-siderophores were synthesized in vitro. Nipponbare rice and its naat1 mutant were reared in soil and gel cultures to determine Fe availability. Hydroponically grown naat1 mutant seedlings were used for reducibility assays to determine the ability of rice roots to reduce Fe(III) chelated by TAGE or DFOB. The expression of a Fe-deficiency inducible gene was also determined, as well as chlorophyll and Fe concentrations. Reduci bility assays on naat1 mutant seedlings revealed that the reduction level of TAGE-Fe(III) was approximately three times higher than that of DFOB-Fe(III). Application of TAGE-Fe(III) to both culture medium and alkaline soil improved Fe chlorosis, growth, and Fe concentration in both naat1 and wild type plants, whereas application of DFOB-Fe(III) only did so in wild type plants. Easily reducible Fe(III)-chelates such as TAGE-Fe(III) can be a better source of Fe for rice plants than most natural microbial siderophores-Fe(III). Our study also demonstrated that rice plants have the ability to utilize microbial siderophores-Fe(III) as the Fe source through the Strategy I-like Fe acquisition system.     

施氮及花后土壤相对含水量对黑粒小麦灌浆期 氮素吸收转运及分配的影响

以黑粒小麦‘漯珍一号’为供试材料,通过棚下盆栽试验研究了不同施氮量及花后土壤相对含水量对‘漯珍一号’植株氮素吸收、转运、分配以及籽粒蛋白质及其组分含量的影响。结果表明:相同施氮量下,黑小麦籽粒含氮量、蛋白质积累量随水分胁迫加剧而降低;各蛋白质组分含量的变化随施氮量的不同而存在差异,在低氮[N_1,150 kg(N)·hm~(-2)]条件下,随水分胁迫加剧,清蛋白、球蛋白、醇溶蛋白含量升高,高氮[N_3,300 kg(N)·hm~(-2)]条件下,清蛋白、球蛋白含量升高,而醇溶蛋白含量降低。相同水分胁迫(土壤相对含水量为55%~65%,W_2;土壤相对含水量为35%~45%,W_3)条件下,籽粒氮素含量、籽粒中蛋白质的积累量随施氮量增加而提高,成熟期籽粒氮素含量占总氮素含量的比例下降;而充足供水(土壤相对含水量为75%~85%,W_1)时,中氮处理[N_2,240 kg(N)·hm~(-2)]籽粒蛋白质积累量最高,同时,营养器官贮藏氮素向籽粒的转运量、转运率均达最大值,对籽粒的贡献率也较高。W_1处理时,清蛋白、球蛋白和醇溶蛋白含量随施氮量的增加而提高,麦谷蛋白在N_2处理达最大值;而W_2、W_3处理情况下,N_2处理小麦中各蛋白质组分含量最高。综上所述,本试验条件下,施氮量及花后土壤相对含水量对黑粒小麦氮代谢具有显著影响,施氮量过高或过低以及水分胁迫均不利于黑粒小麦氮代谢过程的有效进行,综合考虑,花后充足供水(W_1)与中等施氮水平(N_2)组合对黑粒小麦氮素吸收、转运和分配具有较好的调控作用。