大丽轮枝菌拮抗细菌的分离与抗菌物质鉴定

从黄萎病发病棉田未发病棉株根际土样中分离芽孢杆菌,利用对峙培养法对大丽轮枝菌进行平板拮抗实验,筛选出6株对大丽轮枝菌有拮抗作用的芽孢杆菌,其中3株菌发酵代谢产物具有抗菌活性。S4-3菌株发酵液滤液经100℃、30min热处理丧失抗菌活性,发酵液氯仿抽提物不具有抗菌活性,发酵液硫酸铵沉淀物有抑菌活性,但经蛋白酶处理后活性丧失,认为S4-3菌株抑菌活性物质是蛋白质。而菌株S4-5发酵液经100℃热处理后还有较高的抗菌活性,发酵液氯仿抽提物也具有抗菌活性,但硫酸铵沉淀物对病原菌无抑制作用;S5-6菌株发酵液滤液经100℃热处理丧失抗菌活性,硫酸铵沉淀物也不具有抗菌活性,而发酵液氯仿抽提物有抗菌活性,因此认为S4-5和S5-6菌株产生的抑菌物质不是蛋白质类。     

棉花根部拮抗枯萎病菌或黄萎病菌的可培养内生细菌多样性分析

为解析棉花内生细菌的种群结构,更好地开发利用生防内生细菌,以棉花枯萎病菌、黄萎病菌为指示病原真菌,对常抗棉和泗棉3号根部分离获得的内生细菌进行拮抗活性筛选,获得87株至少对其中1个指示病原真菌具有拮抗作用的内生细菌。ARDRA分析(Amplified ribosomal DNA restriction analysis)和16S r DNA序列测定表明棉花根部可培养内生拮抗细菌可分为13个ARDRA类群,分属于芽孢杆菌属(Bacillus)、短芽孢杆菌属(Brevibacillus)、伯克霍尔德氏菌属(Burkholderia)、农杆菌属(Agrobacterium)、中华根瘤菌属(Sinorhizobium)、不动杆菌属(Acinetobacter)和假单孢菌属(Pseudomonas)7个属。其中常抗棉根部内生拮抗细菌分属于芽孢杆菌属、短芽孢杆菌属、伯克霍尔德氏菌属和农杆菌属,而泗棉3号根部内生拮抗细菌则分属于芽孢杆菌属、伯克霍尔德氏菌属、中华根瘤菌属、不动杆菌属和假单孢菌属,二者的优势种群都是芽孢杆菌属细菌。BOX-PCR分析显示这些内生拮抗细菌具有丰富的遗传多样性。通过测定这些内生拮抗细菌对棉花黄萎病菌和棉花枯萎病菌的拮抗力大小,以及是否具有产纤维素酶、几丁质酶、蛋白酶和嗜铁素的活性,表明这些内生拮抗细菌具有不同的生防作用机制。     

棉花黄萎病拮抗蛋白的分离与纯化

从棉花叶组织中分离到的类芽孢杆菌(Paenibacillus)LC105菌株对棉花黄萎病病原菌大丽轮枝菌具有明显的拮抗作用.该菌株发酵液分别用过滤、高压灭菌及有机溶剂处理,初步证明对大丽轮枝菌产生拮抗作用的物质为蛋白质.LC105菌株发酵液经硫酸铵沉淀、DEAE-Sephadex A-50离子交换层析及Sephacryl S-100凝胶层析分离纯化,得到了分子量为27 kDa的抗菌蛋白.     

黄瓜枯萎病生防芽孢杆菌的定向筛选及其抑菌活性物质特性分析

研究拟从北京郊区黄瓜根际土壤中定向筛选黄瓜枯萎病菌生防芽孢杆菌,并研究其抑菌活性物质的基本特性。采用热处理和形态观察结合平板对峙法定向筛选黄瓜枯萎病菌拮抗菌。从北京郊区黄瓜根际土壤中定向分离到63株产芽胞的菌株,从中筛选到2株对黄瓜枯萎病菌（Fusarium oxysporium f.sp.cucumarinum Owen.）具有较好拮抗作用的菌株,命名为j-28和j-50。拮抗菌j-28对黄瓜枯萎病菌的抑菌带达到11.0 mm。此外,拮抗菌j-28有较广的抑菌谱,对黄瓜灰霉病菌（Botrytis cinerea）、马铃薯干腐病菌（Fusarium sulphureum）和茄子黄萎病菌（Verticilllium albo-atrum）等具有很强的拮抗作用。经过形态观察、理化分析和分子鉴定,j-28和j-50鉴定为枯草芽孢杆菌（Bacillus subtilis）。研究还发现拮抗菌j-28的发酵液经过高温和强酸碱处理后抑菌效果会明显减弱,说明拮抗菌j-28产生的主要抑菌活性物质可能是蛋白类。笔者定向筛选到针对北京郊区多发的黄瓜枯萎病菌拮抗菌枯草芽孢杆菌j-28,具有潜在的应用价值,为今后北京郊区黄瓜枯萎病菌的防治提供新的生物防治资源。     

河北省又取得一批农药植保类科研成果

<正>(接上期)7.棉花黄萎病拮抗芽孢菌剂的研制单位名称:河北农业大学评价单位名称:保定市科技局项目组分离1株对棉花黄萎病病原菌大丽轮枝菌(Verticillium dahliae Kleb.)有较高拮抗的芽孢杆菌菌株,经鉴定为枯草芽孢杆菌(Bacillus subtilis)。盆栽试验表明,Z-5菌株对棉花黄萎病的防治效果达到76.05%。采用抗利福平和抗     

棉花黄萎病生防内生芽孢杆菌LH-L3的分离鉴定

【目的】获得能够防治黄萎病菌的棉花内生芽孢杆菌。【方法】将棉株组织研磨液80℃加热后,采用涂板法分离内生菌,将对大丽轮枝菌强致病力菌株Vd084拮抗效果最好的菌株,依据菌体形态、生理生化特征、16S rRNA、gyrB和rpoB基因序列分析进行鉴定,并以盆栽试验确定其对棉花黄萎病的防效。【结果】分离到61株芽孢杆菌类似菌,其中17株能拮抗大丽轮枝菌。复筛得到1株拮抗活性较高、抑菌谱较广的菌株LH-L3。该菌株被鉴定为甲基营养型芽孢杆菌。与无菌水对照相比,用106 mL~(-1)的LH-L3菌液浇灌3次的棉花出苗率、株高、根长、地上以及地下部分鲜物质质量分别提高42.85%、10.24%、23.83%、10.05%、97.62%,对棉花黄萎病的防治效果达到85.24%。【结论】菌株LH-L3有较好的促生长和棉花黄萎病防治的效果,具有进一步开发应用潜力。     

一株大丽轮枝菌拮抗细菌7-47菌株的分离与鉴定

 从全国各地棉田采集土样,筛选对棉花黄萎病致病菌大丽轮枝菌（Verticillium dahliae Kleb.）具有拮抗作用的芽孢杆菌菌株。通过初筛得到具有拮抗作用的菌株68株,复筛选出1株拮抗活性较高的菌株7-47,并通过生理生化试验,形态特征的观察及16S rDNA的序列分析对其进行了鉴定,认为菌株7-47属于漠海威芽孢杆菌（Bacillus mojavensis）。     

生防细菌NCD-2突变体构建及抑菌功能基因的防病作用

生防细菌NCD-2是一株枯草芽孢杆菌菌株,该菌株通过分泌抑菌肽而对棉花黄萎病病原菌和棉花立枯病病原菌起到抑制作用。本研究着重通过原生质体法与诱导转座方法,建立了携带转座子Tn917质粒pTV1对枯草芽孢杆菌NCD-2野生菌株的转化体系与转座子突变技术,获得1500多个转座子插入突变子。通过测定这些突变子对大丽轮枝菌的抑制作用,筛选到2个抗生作用丧失的抑菌功能缺失的突变子。室内盆栽试验结果表明这2个抑菌功能缺失突变子对棉花立枯病的防效显著低于野生菌株,说明NCD-2野生菌株产生的抑菌肽在该菌株防治棉花立枯病中起到主要作用,进而说明编码该抑菌肽的基因在该菌株防治棉花立枯病中具有重要作用。     

金龟子绿僵菌对棉花枯萎病菌的拮抗作用研究

在实验室条件下,研究了虫生真菌金龟子绿僵菌(Metarhizium anisopliae)对棉花枯萎病菌尖孢镰刀菌(Fusarium oxysporium f.sp.vasinfectum)的拮抗作用及其机制。对峙培养结果表明,金龟子绿僵菌的3个供试菌株对3株枯萎病菌均有抑制作用,其中以Ma55抑制效果最好。在培养基中加入Ma55不同浓度的孢子悬浮液,当浓度为3.25×106 cfu.mL-1时,5 d后,对3株枯萎病菌菌丝生长的抑制率均超过70%。Ma55液体振荡培养20 d获得的无菌发酵液对3个枯萎病菌菌株菌丝生长的抑制率分别达到59.86%、56.99%和57.09%,对它们的分生孢子产生及分生孢子萌发也都具有显著的抑制作用。显微镜观察未发现金龟子绿僵菌与棉花枯萎病菌菌丝间有重寄生现象。上述结果显示,金龟子绿僵菌对供试棉花枯萎病菌的拮抗机制主要是营养竞争、空间竞争及抗生作用。     

棉花黄萎病拮抗细菌LC-04菌株的抗菌蛋白产生条件研究

LC-04菌株是从棉花叶部组织中分离出的一株类芽孢杆菌,其发酵液上清液的硫酸铵沉淀物对大丽轮枝菌V-190菌株的生长具有明显的拮抗作用,实验证明发酵液中含有蛋白质类抗菌物质。LC-04菌株的适宜发酵条件为:在pH7的LBG培养基中于30℃,180 r.min-1摇床振荡培养48 h,用70%硫酸铵对LC-04菌株发酵液上清液进行盐析,可以沉淀最多的抗菌蛋白。     

A note on the origin of Kalanchoë x vadensis Boom & Zeilinga (Crassulaceae)

Summary K x vadensis is a hybrid of K. blossfeldiana and K. marmorata obtained after doubling the number of chromosomes.     

Avoidance of leaf rust fungi in wild relatives of cultivated cereals

Summary Avoidance of rust fungi that was based on poor appressorium induction was previously found in Hordeum chilense. In the present study 95 accessions of Triticeae were screened for avoidance of Puccinia hordei. The percentage of appressorium formation per germinated spore ranged from 6 to 90%. On none of the 41 accessions of Aegilops, Agropyron, Elymus, Secale, Thinopyrum or Triticum studied was the rate of appressorium formation lower than 25%. Lower rates of appressorium formation were, however, found on accessions of wild barley species Hordeum brachyantherum, H. marinum, H. parodii and H. secalinum. Its implications in cereal breeding are discussed.     

Relation between wheat-rye crossability and seed set of common wheat after pollination with other species in the Hordeae

Summary Cultivars of common wheat (Triticum aestivum L. em. Thell.) of high wheat-rye (Secale cereale L.) crossability set more seed with pollen of other related species than did wheats of low wheat-rye crossability. This was found to be true for pollen parents from the genera Triticum, Aegilops, Secale, Agropyron and Elymus.     

Mechanisms and diversity of resistance to sorghum shoot fly,Atherigona soccata

Sorghum shoot fly, Atherigona soccata, is one of the important pests of postrainy season sorghums. Of the 90 sorghum genotypes evaluated for resistance to this pest, RHRB 12, ICSV 713, 25026, 93046 and 25027, IS 33844‐5, Giddi Maldandi and RVRT 3 exhibited resistance in postrainy season, while ICSB 463, Phule Anuradha, RHRB 19, Parbhani Moti, ICSV 705, PS 35805, IS 5480, 5622, 17726, 18368 and 34722, RVRT 1, ICSR 93031 and Dagidi Solapur showed resistance in rainy season, suggesting season‐specific expression of resistance to A. soccata. ICSB 461, ICSB 463, Phule Yasodha, M 35‐1, ICSV 700, 711, 25010, 25019 and 93089, IS 18662, Phule Vasudha, IS 18551 and 33844‐5 and Barsizoot had fewer deadhearts than plants with eggs across seasons, suggesting antibiosis as one of the resistance mechanism. Five genotypes exhibited resistance with high grain yield across seasons. Correlation, path and stepwise regression analyses indicated that leaf glossiness, seedling vigour, trichome density, oviposition and leaf sheath pigmentation were associated with the expression of resistance/susceptibility to shoot fly, and these can be used as marker traits to select and develop shoot fly‐resistant sorghums.     

Reaction of tritordeum to Fusarium culmorum and Septoria nodorum

Summary Hordeum chilense is a wild barley extensively used in wide crosses in the Triticeae. It could be a valuable source of resistance to Fusarium culmorum and Septoria nodorum. Some H. chilense x Triticum spp. amphiploids, named tritordeums, were more resistant than the parental wheat line to these diseases, others were not. Average contents of ergosterol and deoxynivalenol (DON) suggested that resistance to colonization by Fusarium was the highest for Hordeum chilense, followed by tritordeum and wheat in decreasing order. In particular, the H. chilense genotypes H7 and H17 enhanced the wheat resistance to F. culmorum in its tritordeum offsprings. Resistance to S. nodorum in tritordeum was not associated with tall plant height. There is sufficient genetic variation for resistance to F. culmorum and S. nodorum among tritordeum to allow the breeding of lines combining short straw and resistance to both diseases.     

Breeding apomictic bahiagrass (Paspalum notatum Flügge) with improved turf traits

The development of a turf‐type bahiagrass could have a remarkable impact on the billion‐dollar turf industry in the south‐eastern United States. The goals of this study were to (i) select bahiagrass genotypes with improved turf attributes and (ii) determine the reproductive mode and seed fertility of selected genotypes. The population included mutants, wild types, hybrids and standard cultivars evaluated at two locations for two years. Morphological variation was observed and clonal repeatability (H²) ranged from 0.27 to 0.90. Flowering was extremely reduced in some mutants, which also exhibited dark green colour, finer texture and reduced canopy height. Cytoembryological observations revealed that diploids reproduced sexually and tetraploids by obligate or facultative apomixis. Genotypes with superior seed fertility were identified, while others were comparable to the standard cultivars. While mutant plants exhibited great morphological variation, the random mutations did not affect their reproductive mode, validating the use of mutagenesis in apomictic species. The high H² indicated that selection for improved turf attributes could be successfully performed with the advantage of clonal seed propagation.     

Disease resistance in protected crops and mushrooms

Summary Cultivars of tomatoes, cucumbers, lettuce and peppers have been bred for resistance to one or more pathogens. Some tomato and cucumber cultivars have resistance to a wide range of diseases. Resistance has been transient in many cases and a succession of cultivars with new genes or new combinations of resistance genes has been necessary to maintain control. There has been a number of notable exceptions and these have included durable resistance to such pathogens asFulvia fulva and tomato mosaic virus. With lettuce the resistance situation is complicated by the occurrence of fungicide resistant pathotypes. There are no strains ofAgaricus bisporus purposely bred for disease resistance.In protected flower crops only resistance to Fusarium wilt in carnations has been purposely bred but differences in disease resistance are apparent in cultivars of many ornamental crops. This is particularly so in chrysanthemums where there are cultivars with resistance to many of the major pathogens. Similar situations occur with other flower crops and pot plants. Cultivars of some species have not been systematically investigated for resistance.The need for genetic resistance will increase with the further reduction, in the limits on pesticide use and an increasing public awareness and importance of pesticide pollution.ADAS is an executive agency of the Ministry of Agiculture, Fisheries and Food and the Welsh Office.     

Inheritance of fruit colour in a wild Russian red raspberry seedling

Summary A new gene, i, from a self-incompatible wild Russian Rubus idaeus seedling is described. The gene i interacts with the fruit colour gene T, Tii plants having apricot instead of red fruits, and spines, leaves and stems pigmented to varying degrees. A deficit of apricot-fruited seedlings in progenies segregating for i could be explained by linkage between i and the self-incompatibility S locus with a crossover value of approximately 26.7%.Segregations for spine colour in crosses and backcrosses of the Russian seedling with two green-spined (ttII) raspberries selected for their yellow fruit colour, showed that both the latter carried the spine colour intensifier gene P. This suggests that earlier authors were incorrect in attributing the apricot fruit colour developed by some tt plants to a pleiotropic effect of P.     

Genetic constitution and diversity in four narrow endemic redwoods from the family Cupressaceae

The genetic constitution and diversity of four relictual redwoods are discussed in this review. These include monotypic genera of the family Cupressaceae: coast redwood (Sequoia sempervirens), giant sequoia (Sequoiadendron giganteum), dawn redwood (Metasequoia glyptostroboides), and alerce (Fitzroya cupressoides). All four species are narrow endemics, share a number of common phenotypic traits, including red wood, and are threatened species. Fossil history suggests that the ancestors of redwoods probably originated during the Cretaceous and Tertiary periods and flourished thereafter for millions of years. Towards the end of the Tertiary period began their decline and struggle for existence that continued during the subsequent geologic upheavals and climate changes, until the survival of the present-day redwoods in the current restricted locations in the world (USA, China, and South America). Although two species, Sequoiadendron and Metasequoia, are diploids (2n = 22), and the other two are polyploids: Fitzroya a tetraploid (2n = 4x = 44), and Sequoia a hexaploid (2n = 6x = 66); they all share the same basic chromosome number x = 11. The genome size in the hexaploid Sequoia is one of the largest (31,500 MB) in the conifers, while the genome sizes of diploid Metasequoia and Sequoiadendron are about one-third (~10,000 MB) of Sequoia. Genetic diversity in the redwoods is lower than most other gymnosperms, except in Sequoia, which seems to rank near the upper quarter of the coniferous forest trees. Genomic research is sparse in the redwoods, and should be pursued for a better understanding of their genome structure, function, and adaptive genetic diversity.     

The induction in vitro of adventitious shoots in Rosa

Summary Adventitious shoots were formed on excised leaves, roots and callus of Rosa persica x xanthina and on excised leaves of R. laevigata and R. wichuraiana on culture media that included BAP and NAA as growth regulators. Shoots formed freely on freshly cultured callus of R. persica x xanthina but their production declined in successive cultures and ceased after twelve weeks. Transplantation to soil was improved by rooting plantlets in cellulose plugs in vitro and transferring plantlets to soil while still in the plugs.