Inhibition of insect pest α-amylases by little and finger millet inhibitors

The inhibitory effect of three proteinaceous inhibitors isolated from little and finger millet was examined on gut α-amylases for four stored grain and four phytophagous insect-pests. Additionally, using native PAGE, several α-amylases isozymes were observed in all insect-pests studied. Furthermore, thermostabilities and the pH optimum for insect-pests α-amylases, which varied from acidic to alkaline, were also determined. On the other hand, proteinaceous inhibitors from little millet seeds Panicum sumatrense (LMCO3) and from finger millet (FMCO11 and FMCO13) inhibited insect-pests α-amylases with different proportions. The highest inhibition percent was recorded for LMCO3 and FMCO13 against Callosobruchus chinensis α-amylase, where the inhibition percent was approximately 70 and 50%, respectively. Furthermore, millet α-amylase inhibitors also reduced significantly digestive α-amylolytic activities of Acaea janata, C. cephalonica, Sitophilus oryzae, and Tribolium castaneum, indicating that these α-amylase inhibitors could be used toward crop insect-pests.     

Resistance to blast (Magnaporthe grisea) in a mini-core collection of finger millet germplasm

Blast caused by Pyricularia grisea [teleomorph: Magnaporthe grisea] is an economically important and widespread disease of finger millet in the world. Host resistance is the most economical and effective means of combating this disease as finger millet is predominantly grown by resource-poor and marginal farmers. At the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), we evaluated a finger millet mini-core collection of 80 germplasm accessions (about 1 % of the total germplasm collection representing major trait variability) for blast resistance both in the field and greenhouse. Field evaluation was done using a refined screening technique that included new improved rating scales for leaf, neck and finger infection. Sixty six of the 80 accessions showed combined resistance to leaf, neck and finger blast in two seasons (2009 and 2010) of field screening. A highly significant and positive correlation was found between neck and finger blast ratings (r?=?0.92), whereas small but significant correlations were found between leaf blast and neck blast (r?=?0.25) and between leaf blast and finger blast (r?=?0.30). These accessions were also screened for leaf blast resistance in the greenhouse by artificial inoculation of seedlings to confirm field observations. Fifty-eight of the 80 accessions were resistant to leaf blast in the greenhouse screen as well. These resistant accessions represented one wild (africana) and four cultivated races (vulgaris, plana, elongate and compacta) of finger millet that originated from 13 countries in Asia and Africa and exhibited considerable diversity for agronomic traits, such as maturity period, plant height and panicle type. These blast resistant accessions from the mini-core collection would be useful in finger millet disease resistance breeding programs.     

Leaf blast disease in finger millet [Eleusine coracana (L.) Gaertn.] caused by Bipolaris sorokiniana Sacc.: a histopathological study

Journal of Plant Diseases and Protection - Finger millet (Eleusine coracana L.) is a staple food in many African and South Asian countries and is ranked fourth among millets produced worldwide....     

谷子抗病基因同源序列的克隆与分析

谷子锈病是谷子上的一种流行性强、毁灭性大的病害,严重影响谷子生产.种植抗病品种是防治锈病最经济有效的方法,但谷子抗锈病种质资源非 常贫乏,而且高抗锈病的材料其农艺性状又很差.很难通过传统育种方法培育抗锈病品种,因此克隆谷子抗锈病基因尤为重要.目前克隆到的许多植 物抗病基因编码的氨基酸序列都有一定的保守结构域.根据抗病基因保守结构域,已克隆的抗病基因主要分为5类,其中最主要的是NBS-LRR(nu- leotide-binding site leucine-rich epeat)和STK(Se-rine／Threonine protein kinase)类.因而根据抗病基因保守结构域设计引物,从植物的DNA中扩增植物的抗病基因同源序列RGA(resistance geneanalogs)更加快捷有效,目前通过RGA方法克隆植物抗病基因已有报道[1,2].     

不同生育阶段灌水处理对谷子生长发育的调控效应

在敦煌年降雨量39 mm条件下,研究了拔节期、抽穗期、灌浆期不同生育阶段灌水处理对谷子农艺性状及产量的影响。试验结果表明:不同生育阶段灌水对谷子主要农艺性状及产量都有较大影响,与全生育期不灌水相比,拔节期、抽穗期、灌浆期各灌1次水处理谷子生育期缩短5.4 d,株高增高51.1 cm,穗长增长1.2 cm,株穗重、株粒重和株草重分别增重14.2、12.3 g和3.1 g,产量增加225.85 kg·666.7m-2;全生育期不灌水处理谷子产量最低,仅105.78 kg·666.7m-2,抽穗期和灌浆期共灌2次水,拔节期、抽穗期、灌浆期共灌3次水和抽穗期仅灌1次水各处理,产量分别达339.91、331.64 kg·666.7m-2和288.31 kg·666.7m-2,灌2次水产量最高。表明抽穗期灌水对谷子产量形成影响最为显著。灰色关联度分析表明,单株粒重、单株穗重和主穗长与产量相关性最为密切。     

Influence of temperature on plant–rhizobacteria interactions related to biocontrol potential for suppression of fusarium wilt of chickpea

Seed and soil treatment with Pseudomonas fluorescens RGAF 19, P. fluorescens RG 26, Bacillus megaterium RGAF 51 and Paenibacillus macerans RGAF 101 can suppress fusarium wilt of chickpea ( Cicer arietinum ), but the extent of disease suppression by these rhizobacteria is modulated by soil temperature. In this work, the effect of temperature on plant–rhizobacteria interactions was assessed in relation to biocontrol potential for suppression of fusarium wilt of chickpea. Seed and soil treatment with those rhizobacteria delayed seedling emergence compared with nontreated controls, and either increased or had no deleterious effect on chickpea growth. Pseudomonas fluorescens isolates significantly increased chickpea shoot dry weight at 20°C and root dry weight at 25 and 30°C. All bacterial isolates colonized the chickpea rhizosphere and internal stem tissues at 20, 25 and 30°C, and there was a positive linear trend between bacterial population size in the rhizosphere and temperature increase. The maximum inhibition of mycelial growth and conidial germination of Fusarium oxysporum f. sp. ciceris race 5 in vitro occurred at a temperature range optimal for bacterial growth and production of inhibitory metabolites. These results demonstrate the need to understand the effects of environmental factors on the biological activities of introduced rhizobacteria of significant importance for plant disease suppression.     

Screening pearl millet cultivars by ELISA for resistance to downy mildew disease

In an earlier study, we described identification of a protein from a virulent pathotype of Sclerospora graminicola , the binding reaction of which differentiated susceptible and resistant cultivars of pearl millet to downy mildew disease. This protein and corresponding antibody were used in an enzyme-linked immunosorbent assay (ELISA) to screen suspension cells of pearl millet cultivars for their resistance to the downy mildew pathogen. Screening results for 31 pearl millet cultivars correlated positively with the established field screening method.     

蔬菜作物根际促生菌分离筛选、鉴定及促生特性测定

为从4种蔬菜作物根际分离和筛选植物根际促生菌(plant growth promoting rhizobacteria,PGPR)并研究其促生特性,采用固氮、无机和有机磷培养基筛选根际促生菌株,对其促生特性进行定性和定量分析,并通过形态特征及16SrDNA序列分析对优良促生菌株进行鉴定。结果表明:从4种蔬菜根际中共获得57株PGPR菌株,其中固氮菌24株,溶磷菌33株。筛选出的19株优良PGPR菌株的固氮酶活性在0.14~3 664.97 nmol C2H4·h~(-1)·mL~(-1)之间,以NCRS1菌株固氮酶活性最高;溶解无机磷菌株的溶磷量在36.99~362.60μg/mL之间,培养液p H为4.57~5.75,以NCRP2菌株溶磷量最大;溶解有机磷菌株的溶磷量在9.24~55.21μg/mL之间,培养液p H为6.24~7.75,以PPRS3菌株溶磷量最大;菌株分泌IAA量均小于3.36μg/mL;NCRP2菌株对小麦长蠕孢病菌Helminthosporium tritici-vulgaris、番茄早疫病菌Alternaria solani、黄瓜枯萎病菌Fusarium oxysporum、马铃薯立枯丝核病菌Rhizoctonia solani、油菜菌核病菌Sclerotinias clerotiorum和玉米小斑病菌Bipolaria maydis均有抑制作用。对19株优良PGPR菌株进行初步鉴定,分属于假单胞菌属Pseudomonas、不动细菌属Acinetobacter、Advenella、叶杆菌属Phyllobacterium、细杆菌属Microbacterium、芽胞杆菌属Bacillus。本研究筛选出的优良PGPR菌株,可为今后生物菌肥推广应用提供菌种资源。     

种植密度对糜子生长发育及产量影响

采用完全随机区组试验设计,以河曲大红黍为材料,设定6个密度处理,研究不同种植密度对糜子的株高、茎粗、分蘖数等形态指标,单株叶面积、叶绿素相对值、光合速率等光合指标,干物质积累量,产量构成因素及产量的影响;运用非线性回归分析的方法,对密度与作物产量的经验模型进行模拟.结果表明:种植密度超过一定范围值时,随着密度的增加,株高增大,茎粗变细,分蘖数减少和光合生理指标均降低,穗重、穗粒重和有效穗粒数逐渐减少,干物质积累量减少,产量降低;产量随着种植密度增加呈先升后降的趋势,二次曲线模型的方程式为Y=176.78471+ 11.51879X-1.29607X2,理论最适密度范围为4万株/666.7m2-5万株/666.7m2.     

生物炭对谷子幼苗生长及光合特性的影响

为揭示生物炭在谷子种植中的应用潜力和价值,以祥谷3号为材料,采用室内盆栽试验的方式,在土壤中添加3种不同比例的生物炭(10、50、90 g·kg~(-1)),于播种后30 d对谷子幼苗进行生物量、根系形态和地上形态及叶片光合的测定分析。结果表明:与未施用生物炭(BC0)对照相比,不同剂量的生物炭均显著改善了谷子幼苗地上和地下生物量、形态指标以及除胞间CO_2浓度以外的其他光合参数指标,且以中等剂量生物炭(50 g·kg~(-1))对谷子生长和光合指标的影响效果最为明显。与对照相比,中等剂量生物炭(50 g·kg~(-1))处理谷子幼苗地上部分和地下部分鲜重、干重分别提高了220%、181%和500%、350%,根冠比增加41.7%;谷子幼苗地下部分主根长、总根长、总根表面积、根尖数、分枝数分别提高124%、215%、323%、86.4%、279%;谷子地上部分幼苗株高、茎粗和叶面积分别提高48.28%、50%、159%;叶片相对叶绿素含量和净光合速率分别提高21.01%和167%。研究表明,生物炭添加可以改善地下根系和地上茎叶形态,改善叶片光合和根系对水分和矿质元素的吸收能力,从而促进谷子生物量的积累,可为后期产量的形成奠定基础。     

盐碱胁迫下一株促进苜蓿生长的细菌筛选与鉴定

为改良和利用盐碱地,本研究以大庆草原盐碱土为材料,在Na₂CO₃和NaHCO₃终浓度分别为50、100 mmol·L^-1,pH 8.5的条件下对其中耐盐碱细菌进行分离,并在CAS铁载体检测培养基、ADF培养基、PKO无机磷培养基等功能性培养基上对菌株的促生功能进行检测;利用16S rDNA序列分析法进行菌株的菌种鉴定,在pH 8.5,25 mmol·L^-1 Na₂CO₃胁迫下测试菌株对苜蓿种子发芽率、发芽势及幼苗生长的影响。结果表明：筛选得到的菌株S11具有产植物激素IAA和产ACC脱氨酶功能;通过对菌株16S rDNA序列进行分析,初步鉴定菌株S11为堀越氏芽孢杆菌（Bacillus horikoshii）。盐碱胁迫条件下,浸种接菌处理苜蓿种子后,种子发芽率和发芽势分别提高了15%和10%,幼苗根长、株高、鲜重以及叶绿素含量分别增加了34.78%、12.5%、27.3%和43.30%,这证明菌株S11具有缓解盐碱胁迫促进苜蓿生长的功能。     

水生拉恩氏菌ZF7分离鉴定及其促生防病效果研究

 从樱花根际土壤中分离到一株对大白菜黑腐病菌具有强拮抗作用的细菌ZF7。根据生物学特征、16S rDNA 序列分析、Biolog及生理生化分析,菌株ZF7被鉴定为水生拉恩氏菌Rahnella aquatilis。利用平板对峙法和双层培养法,证明菌株ZF7具有较广的抑菌谱,对5种病原真菌和5种病原细菌具有明显拮抗效果。而且,该菌能在大白菜根际土壤以及白菜根部稳定定殖。采用盆栽试验对菌株ZF7进行促生防病效果测定,结果表明该菌对大白菜具有显著的促生效果。灌根菌量为20 mL/株时,菌株ZF7对大白菜株高和根长的促生率分别达56.47%、54.74%,对地上部重量及根重的促生率分别达233.84%和164.71%。同时,菌株ZF7对大白菜黑腐病具有良好的防治效果,防效可达88.80%,高出中生菌素的防治效果26.76%。综上分析,菌株ZF7生防性状优良,具有一定的田间应用开发潜力。     

水生拉恩氏菌ZF7分离鉴定及其促生防病效果研究

 从樱花根际土壤中分离到一株对大白菜黑腐病菌具有强拮抗作用的细菌ZF7。根据生物学特征、16S rDNA 序列分析、Biolog及生理生化分析,菌株ZF7被鉴定为水生拉恩氏菌Rahnella aquatilis。利用平板对峙法和双层培养法,证明菌株ZF7具有较广的抑菌谱,对5种病原真菌和5种病原细菌具有明显拮抗效果。而且,该菌能在大白菜根际土壤以及白菜根部稳定定殖。采用盆栽试验对菌株ZF7进行促生防病效果测定,结果表明该菌对大白菜具有显著的促生效果。灌根菌量为20 mL/株时,菌株ZF7对大白菜株高和根长的促生率分别达56.47%、54.74%,对地上部重量及根重的促生率分别达233.84%和164.71%。同时,菌株ZF7对大白菜黑腐病具有良好的防治效果,防效可达88.80%,高出中生菌素的防治效果26.76%。综上分析,菌株ZF7生防性状优良,具有一定的田间应用开发潜力。     

Cellulysin induces downy mildew disease resistance in pearl millet driven through defense response

The responses to cellulysin as an immune inducer in pearl millet that confers downy mildew resistance mediated through lipoxygenase (LOX), a jasmonate-dependent enzyme involved in defence signalling, are discussed in this paper. The susceptible pearl millet cultivar 7042S was treated with cellulysin at 10, 15, 20, 30 and 50 μg/ml concentrations. All tested concentrations showed enhanced seed germination and seedling vigour when compared with the untreated control. Maximum seed germination of 92 % and seedling vigour was obtained following 20 μg/ml cellulysin treatment. Significant (P?<?0.05) downy mildew disease protection of 67 % and 71 % was observed when cellulysin was used at 20 μg/ml under greenhouse and field conditions, respectively. Further studies showed that the resistance induced by cellulysin treatment in pearl millet plant was systemic, required a minimum of 4 days to achieve maximum resistance development after pathogen inoculation seedling inoculation (five-day-old), and was sustained throughout the plant’s life. Plants raised from cellulysin-treated seeds and challenge inoculated at tillering (25-day-old) and inflorescence (45-day-old) showed persistence in resistance till the end of the crop period. A notable increase in LOX activity was observed in all the tested concentrations of cellulysin in plants inoculated with the pathogen at 24 h, compared to the control. However, a maximum 6-fold increase in LOX activity was noticed using a cellulysin concentration of 20 μg/ml 48 hours post inoculation. In contrast, glucanase (GLU) activity was high in control inoculated seedlings, but was low in cellulysin treated samples at all time intervals. The optimal cellulysin treatment (20 μg/ml) provided enhanced vegetative and reproductive parameters that resulted in higher yield compared to the untreated control.     

AMF+PGPR组合提高黄瓜抗枯萎病的作用机制

对黄瓜(Cucumis sativus)品种香翠16接种尖孢镰刀菌黄瓜专化型(Fusarium oxysporum f.sp.cucumerinum,Foc)、丛枝菌根真菌(AMF)Funneliformis mosseae(Fm)+Foc、丛枝菌根真菌Glomus versiforme(Gv)+Foc、根围促生细菌(PGPR)PS2-6+Foc、PS3-2+Foc、Fm+PS3-2+Foc以及Gv+PS2-6+Foc,以不接种处理作为对照。结果表明,接种Foc条件下,Fm+PS3-2和Gv+PS2-6组合处理的抗病效果显著优于单接种AMF或PGPR处理;镜检观测到靠近AMF菌丝的Foc菌丝纤细,生长不良。AMF菌丝能缠绕Foc菌丝,并阻挡其侵入根系;AMF侵染率与其防效均呈正相关(P0.05);Fm+PS3-2+Foc和Gv+PS2-6+Foc处理的根系活力是单接种Foc的4.6倍;与不接种对照相比,Fm+PS3-2+Foc和Gv+PS2-6+Foc处理的黄瓜叶片中吲哚乙酸(IAA)含量分别提高了205%和166%,赤霉素(GA3)提高了33%和22%,玉米素核甘(ZR)提高了100%和80%,JA平均提高了65%,ABA降低23%和17%;而与单接种Foc处理相比,IAA提高了667%和568%,GA3提高了223%和196%,ZR提高了254%和219%,茉莉酸(JA)含量平均提高了215%,而脱落酸(ABA)含量只有单接种Foc处理的53%;Fm+PS3-2+Foc和Gv+PS2-6+Foc处理显著提高了黄瓜叶内苯丙氨酸解氨酶(PAL)防御酶活性和PAL基因表达量。综上推断AMF+PGPR组合是通过拮抗病原物、促进合成抗病信号物质、上调防御基因表达、提高防御酶活性和降低植株体内有毒物质等机制降低病害和改善植物抗病性。     

植物根际优良促生菌（PGPR）筛选及其接种剂部分替代化肥对玉米生长影响研究

通过测定植物根际促生菌株固氮酶活性、溶磷量、分泌植物生长素、对病原菌的拮抗作用及菌株生长速度,筛选出了5株具有较好促生特性的优良菌株.利用筛选的优良菌株研制复合接种剂,并于2009年和2010年进行田试,测定其部分替代化肥对玉米生长的影响.结果表明:利用筛选的优良促生菌研制的复合微生物接种剂符合《微生物肥料》NY227-94标准.施用接种剂替代20％ ～ 30％的化肥,玉米株高、地上植物量、穗长、穗粗、单位面积穗数、穗粒数和经济产量等均有提高.研制的微生物接种剂(菌肥+80％化肥),2010年大田推广使用,玉米增产9.86％,减少化肥投入、增产收入及直接经济效益分别为620.1元,hm-2、2 291.4元,hm-2和2 851.5元.hm-2.     

Predictive factors for the suppression of fusarium wilt of tomato in plant growth media

ABSTRACT Fusarium wilts are economically important diseases for which there are no effective chemical control measures. However, biological control and fertility management are becoming efficient alternatives for controlling this disease. Growth media formulated with composts that are able to suppress Fusarium wilt of tomato provide a control system that integrates both strategies. The aim of this study was to predict Fusarium wilt suppression of growth media using abiotic and biotic variables. Grape marc compost was the most effective medium used to suppress Fusarium wilt. Cork compost was intermediate, and light peat and expanded vermiculite were the most conducive growth media. The growth media evaluated were in a pH range of 6.26 to 7.97. Both composts had high beta-glucosidase activity. When pH and beta-glucosidase activity were taken into account as predictive variables, more than 91% of the variation in severity of Fusarium wilt was explained. This relationship illustrates the effect of nutrient availability and the degree of microbiostasis, two key factors in this pathosystem. Microbial populations involved in suppressiveness were cellulolytic and oligotrophic actinomycetes, fungi, and the ratios cellulolytic actinomycetes/cellulolytic bacteria, oligotrophic bacteria/copiotrophic bacteria, and oligotrophic actinomycetes/oligotrophic bacteria. Based on community level physiological profiles, different community structures were evident among growth media evaluated.     

切断深层根对黍子根系及地上部营养生长的影响

采用根管土柱栽培的方法,研究了拔节期和抽穗期切断不同深度(20 cm和40 cm)根系对黍子根系整体及地上部营养生长的影响。结果表明,与不断根相比,在抽穗期和拔节期断掉20 cm或40 cm以下根系,均能导致黍子株高、旗叶叶绿素含量、旗叶SOD与POD活性、单株绿叶面积、总根活力、总根长、总根重及产量明显降低,旗叶M DA含量明显增高。抽穗期断深层根对黍子的影响大于拔节期。但同一生育时期不同深度断根处理间黍子产量差异未达显著水平,表明深层根系(40 cm以下根系)对产量的贡献更大。     

Spore cell wall components ofAspergillus niger elicit downy mildew disease resistance in pearl millet

Elicitors derived from the cell wall of fungi are shown to be active in eliciting resistance in plants against a wide range of pathogens. In the present study carbohydrate components from the autoclaved spore cell wall ofAspergillus niger were prepared as aqueous suspensions and tested for defense response in pearl millet (Pennisetum glaucum (L.) R.Br.) against the oomycetous downy mildew pathogenSclerospora graminicola (Sacc.) Schroet. The aqueous suspension derived from the spore cell wall ofA. niger was used as a seed soak treatment at concentrations of 0.25, 0.5, 1.0, 1.5 and 2.0 mg ml⁻¹ for time intervals of 3, 6, 9 and 12 h. The concentration of 0.5 mg ml⁻¹ for a 6 h soaking period offered 94% seed germination and seedling vigor index increased to 1526. The seed germination and the seedling vigor were significantly higher than the untreated check. Spore cell wall suspension as seed treatment at a concentration of 0.5 mg ml⁻¹ required a 3-day time interval to provide 67% protection against downy mildew. Histological and biochemical studies were conducted to elucidate the mechanism of defense response in treated seedlings uponS. graminicola infection. Resistance host response was detected in the form of lignin and callose deposition in the epidermal cell wall of pearl millet seedlings, which is the site ofS. graminicola infection. A time course study showed rapid and localized deposition of lignin and callose in epidermal cell wall of carbohydrate components-treated pearl millet seedling coleoptiles. Increased levels of the defense-related enzyme peroxidase were detected in the treated seedlings. Peroxidase activity in elicitor-treated samples reached a peak at 8 h post-infection, which was 45% more than in their respective uninoculated control. Characterization of peroxidase isoforms by isoelectric focusing revealed 16 different isoforms, of which pI 6.8, 7.2 and 8.7 increased in elicitor-treated samples uponS. graminicola infection. http://www.phytoparasitica.org posting Nov. 14, 2005.