大豆早熟耐病品种对大豆胞囊线虫耐病机制的初步研究

经2012年和2013年大田试验筛选,选择对大豆胞囊线虫具有耐病性的大豆早熟品种Красноградская,以感病品种黑河52作对照进行了耐病机制初步研究。结果表明,室内人工接种大豆胞囊线虫,接种后6、12、18、24、30 d,大豆耐病品种和感病品种植株根内多酚氧化酶(PPO)活性均略高于未接种的对照,但差异不显著;受大豆胞囊线虫感染,耐、感大豆品种植株根内过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)活性都比同期的对照显著升高,且耐病品种POD活性始终高于感病品种,活性增高维持的时间比感病品种长。同时,PAL活性比感病品种高峰值出现的早。叶绿素含量变化结果显示,大豆胞囊线虫的侵染显著影响其光合作用,但随接种时间的延长,品种间变化规律不同。在接种后的12~24 d,无论耐、感品种叶绿素的含量与各自的对照相比都有所升高,而且耐病品种的上升幅度明显高于感病品种,24 d以后,感病品种的叶绿素含量开始下降,而耐病品种则继续维持在一个较高的水平。     

拟粗壮螺旋线虫对棉花的致病力及其与棉枯萎病的复合症

 温室测定结果:拟粗壮螺旋线虫(Helicotylenchus pseudorobusius)接种量在每100cm³土壤100~1000条的条件下,接种30天后棉苗生长量比无线虫对照苗显著降低(P<0.05)。抗枯萎品种86-1在单接枯萎菌无线虫处理中不发生枯萎病,在枯萎菌(Fusarium oxysporum f. sp. vasinfectum)接种量为每克土7.5×10⁵孢子和线虫(每100cm³±500~1000条)组合处理中发病,棉苗枯萎病情指数与线虫接种量之间呈正相关(γ=0.97)。田间试验结果表明:防治线虫和枯萎病复合症用薰蒸剂棉隆每m²8~14g处理土壤比用非薰蒸性杀线虫剂克百威(呋哺丹),丙线磷(益收宝)和克线磷的效果更好。     

巧防果树再植病

在农业产业结构调整中 ,果树品种的更新换代和矮化密植栽培的发展 ,常常因重复栽植而导致果树发病 ,果农称之为果树再植病 ,其表现为 :果树生长发育受阻 ,产量、品质下降、成活率低、寿命短、甚至出现早衰现象。引起果树再植病的原因很多 ,但主要是由于某些元素的过度缺乏 (或过剩 ) ,土壤中残留毒害物质等引起的 ,大多数果树均能发生再植病 ,但以苹果和桃树表现最为严重。根据笔者多年实践 ,防治果树再植病的方法有以下几种 :1　换树种栽植各树种间前后批再植病的发病情况 :1.1　前批为桃树 ,后批栽杏、梨则不会发生再植病 ,而种植葡萄、桃…     

不同大豆品种与SCN 3号生理小种互作效应研究

 在田间研究了不同大豆品种对大豆胞囊线虫(soybean cyst nematode,SCN)的发育及繁殖的影响不同。结果表明,辽K89102为抗侵入型品种,Peking、PI90763、应县小黑豆、磨石豆和Franklin等的根内线虫从J2向J3或J3向J4发育过程中受到抑制,这些品种均有不同程度的抗线虫发育特性;不同品种对其根内SCN的雌雄分化有较大影响,感病品种根内SCN雌雄比远大于抗病品种,辽豆10根内线虫雌雄比例最高,为11.5:1.0,Peking根内线虫雌雄比例最低,为0.1:1.0;不同品种根围SCN最终群体量有较大差异,辽豆10根围土壤中胞囊量增加了331.5%,Peking和磨石豆的增幅最小,为16.7%。SCN侵染后,不同品种产生的组织病理学反应不同,抗病品种产生明显的组织病理学反应,表现在品种Peking和PI437654的根内线虫虫体附近根组织细胞有坏死现象,而感病品种辽豆10和开育10则没有坏死现象。     

品种、培养基质和接种量对马铃薯脱毒基础苗生长的影响

以马铃薯早熟品种大西洋、中熟品种克新1号、晚熟品种青薯168脱毒基础苗为试验材料,研究品种、培养基质和接种量对马铃薯脱毒基础苗苗鲜重、根鲜重、苗干重、根干重、叶片数、株高、茎粗、根长和根条数等性状指标的影响,培养14 d时,测定各性状指标。结果表明:三个品种的主要农艺性状均在液体培养基上表现为最佳,其平均单株鲜重、干重、株高、茎粗、节长和根条数分别为:159.4 mg、9.1 mg、7.3 cm、1.16 mm、1.4 cm、5.6条。在液体培养基上,降低了马铃薯工厂化生产成本,同时还可缩短马铃薯脱毒基础苗培养时间7~9 d。接种量对马铃薯单株根鲜重、苗干重、叶片数、株高、茎粗、根长有显著性影响(P<0.05);不同成熟期品种苗鲜重、根鲜重、根干重、叶片数、株高、茎粗、根长和根条数受品种影响显著(P<0.05);不同成熟期品种、不同接种密度处理对马铃薯单株根鲜重、苗干重、根干重、叶片数、株高、茎粗、根长影响有显著性交互作用(P<0.05);早熟品种大西洋,接种密度为0.67株/cm2时,中熟品种克新1号和晚熟品种青薯168接种密度为0.80株/cm2时,综合农艺性状表现最好。     

施用生防菌菌剂与日光消毒和生物熏蒸相结合防治番茄根结线虫的研究

在温室条件下,研究了覆膜日光消毒、有机物料生物熏蒸和接种根结线虫拮抗菌Bacillus cereus X5对番茄生物量、根结线虫侵染以及根际土壤和土体土壤中微生物数量变化的影响。结果表明,对根结线虫滋生土壤进行覆膜日光消毒、生物熏蒸和接种拮抗菌的联合处理效果最佳,优于单一处理。联合处理的番茄生物量干重比对照增加了1.8倍;每株番茄根结线虫卵块数和每卵块的虫卵数量分别减少了190.4%和380.5%,对根结线虫的防治效果达到82.7%;根际土壤和土体土壤的细菌和真菌数量显著增加;土壤中根结线虫的数量显著减少。     

松材线虫萎蔫病在我国南方林区的流行能力

通过人工接种松材线虫测定6个“北带”马尾松种源和2个“中带”马尾松种源的3～6年生苗的感病性。接种苗枯死率为37.5%至80%,与马尾松在全国现有疫区内普遍不同程度自然发病枯死的情形一致。7个种源接种苗的平均枯死率(53.5%)低于黑松对照(89.4%)。表明国内的马尾松种源对松材线虫中等或中等以上感病。测定国内重要疫区的5个代表性松材线虫株对8年生黑松和3年生马尾松苗的致病力,枯株率分别为90%以上和30%～90%。由此分析了松材线虫萎蔫病在南方林区流行的极大可能性。     

生姜根结线虫病原鉴定及发生规律

采用田间调查、接种试验、电镜与显微镜观察以及酯酶同工酶电泳等方法,对引发生姜癞皮病的病原及发生规律进行了研究.结果表明,引起生姜癞皮病的病原为南方根结线虫Meloidogyne incognita.该病在每年6月中旬开始发生,8、9月份危害严重.病原在生姜上一年可发生完整的4代,完成1代平均约需35天.病原主要在0～40cm的土层内分布和危害,但具体分布情况依寄主生长状况而稍有差异.南方根结线虫繁殖速率受初始接种密度的影响也很大,当初始接种密度较低时,线虫繁殖速率较高,初始接种密度增大,繁殖速率降低,其平衡密度为每100g干土746.20个卵.     

松材线虫伴生细菌与松树萎蔫病关系的初步研究

利用马尾松水培离体松枝作接种材料,接种消毒后的松材线虫、线虫虫体上分离到细菌分离物GD1以及两者的混合体。结果表明,单独接种消毒后的松材线虫蔌细菌分离物GD1均不能使松枝萎蔫,只有两者混合接种才能使松枝发病枯死。从而说明了松材线虫的伴生细菌对松树萎蔫病的发生起着重要的作用。     

水稻潜根线虫病田间消长与产量损失研究

经对水稻多品种定田定期取定量稻根进行分离观察,结果表明:早晚稻潜根线虫(Hirschmanniella spp.)分别在水稻破口期和齐穗期各有一个高峰。采用盆栽接种试验表明:接种量越大,产量越低,呈显著负相关;分蘖盛期前接种减产显著。该线虫主要影响千粒重和有效穗数。     

Impact of wheat cultivation on microbial communities from replant soils and apple growth in greenhouse trials

ABSTRACT Studies were conducted to assess the impact of short-term rotations of wheat on microbial community composition and growth of apple in soils from replant orchard sites. Soils from two orchards were cultivated with three successive 28-day growth cycles of 'Eltan', 'Penewawa', or 'Rely' wheat in the greenhouse and subsequently planted to 'Gala' apple seedlings. Cultivation of orchard replant soils with any of the three wheat cultivars enhanced growth of apple relative to that achieved in untreated soils. Improved growth was associated with a marked reduction in apple root infection by species of Rhizoctonia and Pythium. Populations of plant-parasitic nematodes were below damage threshold levels in these orchard soils; however, apple seedlings grown in wheat-cultivated soils had significantly lower root populations of Pratylenchus spp. than did seedlings grown in untreated soils. Growth of apple in 'Penewawa'-cultivated soils often was superior to that observed in soils planted with 'Eltan' or 'Rely'. In untreated orchard soils, fluorescent pseudomonad populations isolated from soil and the apple rhizosphere were dominated by Pseudomonas fluorescens biotype C and Pseudomonas syringae. Cultivation of replant soils with wheat induced a characteristic transformation of the fluorescent pseudomonad population, and Pseudomonas putida dominated the population of this bacterial group recovered from wheat-cultivated replant orchard soils. Results from this study suggest that use of short-term wheat cropping sequences during orchard renovation could be useful in management of replant disease and that this disease-control option may operate, in part, through modification of the fluorescent pseudomonad community.     

Suppression of Specific Apple Root Pathogens by Brassica napus Seed Meal Amendment Regardless of Glucosinolate Content

ABSTRACT The impact of Brassica napus seed meal on the microbial complex that incites apple replant disease was evaluated in greenhouse trials. Regardless of glucosinolate content, seed meal amendment at a rate of 0.1% (vol/vol) significantly enhanced growth of apple and suppressed apple root infection by Rhizoctonia spp. and Pratylenchus penetrans. High glucosinolate B. napus cv. Dwarf Essex seed meal amendments did not consistently suppress soil populations of Pythium spp. or apple root infection by this pathogen. Application of a low glucosinolate containing B. napus seed meal at a rate of 1.0% (vol/vol) resulted in a significant increase in recovery of Pythium spp. from apple roots, and a corresponding reduction in apple seedling root biomass. When applied at lower rates, B. napus seed meal amendments enhanced populations of fluorescent Pseudomonas spp., but these bacteria were not recovered from soils amended with seed meal at a rate of 2% (vol/vol). Seed meal amendments resulted in increased soil populations of total bacteria and actinomycetes. B. napus cv. Dwarf Essex seed meal amendments were phytotoxic to apple when applied at a rate of 2% (vol/vol), and phytotoxicity was not diminished when planting was delayed for as long as 12 weeks after application. These findings suggest that B. napus seed meal amendments can be a useful tool in the management of apple replant disease and, in the case of Rhizoctonia spp., that disease control operates through mechanisms other than production of glucosinolate hydrolysis products.     

Elucidation of the microbial complex having a causal role in the development of apple replant disease in washington

ABSTRACT Systematic studies were conducted to elucidate the role of different soil microbial groups in the development of apple replant disease. Populations of targeted microorganisms were reduced by the application of semiselective biocides and soil pasteurization. Bacteria were not implicated in the disease, because application of the antibiotic chloramphenicol reduced soil populations of bacteria but failed to improve growth of apple transplants, while enhanced growth was achieved at pasteurization temperatures that did not alter attributes of the bacterial community recovered from apple roots. Populations of Pratylenchus penetrans were below the damage threshold level in eight of nine orchards surveyed, and nematicide applications failed to enhance apple growth in four of five replant soils tested, indicating that plant parasitic nematodes have a minor role or no role in disease development. Application of the fungicide difenconazole or metalaxyl enhanced growth of apple in all five replant soils, as did fludioxinil in the two soils tested. Soil pasteurization enhanced growth of apple and resulted in specific changes in the composition of the fungal community isolated from the roots of apple seedlings grown in these treated soils. Cylindrocarpon destructans, Phytophthora cactorum, Pythium spp., and Rhizoctonia solani were consistently isolated from symptomatic trees in the field and were pathogenic to apple. However, the composition of the Pythium and Rhizoctonia component and the relative contribution of any one component of this fungal complex to disease development varied among the study orchards. These findings clearly demonstrate that fungi are the dominant causal agents of apple replant disease in Washington state.     

Cultural management of microbial community structure to enhance growth of apple in replant soils

ABSTRACT Apple replant disease typically is managed through pre-plant application of broad-spectrum soil fumigants including methyl bromide. The impending loss or restricted use of soil fumigants and the needs of an expanding organic tree fruit industry necessitate the development of alternative control measures. The microbial community resident in a wheat field soil was shown to suppress components of the microbial complex that incites apple replant disease. Pseudomonas putida was the primary fluorescent pseudomonad recovered from suppressive soil, whereas Pseudomonas fluorescens bv. III was dominant in a conducive soil; the latter developed within 3 years of orchard establishment at the same site. In greenhouse studies, cultivation of wheat in replant orchard soils prior to planting apple suppressed disease development. Disease suppression was induced in a wheat cultivar-specific manner. Wheat cultivars that enhanced apple seedling growth altered the dominant fluorescent pseudo-monad from Pseudomonas fluorescens bv. III to Pseudomonas putida. The microbial community resident in replant orchard soils after growing wheat also was suppressive to an introduced isolate of Rhizoctonia solani anastomosis group 5, which causes root rot of apple. Incorporation of high glucosinolate containing rapeseed ('Dwarf Essex') meal also enhanced growth of apple in replant soils through suppression of Rhizoc-tonia spp., Cylindrocarpon spp., and Pratylenchus penetrans. Integration of these methods will require knowledge of the impact of the biofumigant component on the wheat-induced disease-suppressive microbial community. Implementation of these control strategies for management of apple replant disease awaits confirmation from ongoing field validation trials.     

Mechanism of action and efficacy of seed meal-induced pathogen suppression differ in a brassicaceae species and time-dependent manner

ABSTRACT The effect of seed meals derived from Brassica juncea, B. napus, or Sinapis alba on suppression of soilborne pathogens inciting replant disease of apple was evaluated in greenhouse trials. Regardless of plant source, seed meal amendment significantly improved apple growth in all orchard soils; however, relative differences in pathogen suppression were observed. All seed meals suppressed root infection by native Rhizoctonia spp. and an introduced isolate of Rhizoctonia solani AG-5, though B. juncea seed meal often generated a lower level of disease control relative to other seed meal types. When introduction of the pathogen was delayed until 4 to 8 weeks post seed meal amendment, disease suppression was associated with proliferation of resident Streptomyces spp. and not qualitative or quantitative attributes of seed meal glucosinolate content. Using the same experimental system, when soils were pasteurized prior to pathogen infestation, control of R. solani was eliminated regardless of seed meal type. In the case of B. juncea seed meal amendment, the mechanism of R. solani suppression varied in a temporal manner, which initially was associated with the generation of allylisothiocyanate and was not affected by soil pasteurization. Among those tested, only B. juncea seed meal did not stimulate orchard soil populations of Pythium spp. and infection of apple roots by these oomycetes. Although application of B. napus seed meal alone consistently induced an increase in Pythium spp. populations, no significant increase in Pythium spp. populations was observed in response to a composite B. juncea and B. napus seed meal amendment. Suppression of soil populations and root infestation by Pratylenchus spp. was dependent upon seed meal type, with only B. juncea providing sustained nematode control. Collectively, these studies suggest that use of a composite B. juncea and B. napus seed meal mixture can provide superior control of the pathogen complex inciting apple replant disease relative to either seed meal used alone.     

Identification and Pathogenicity of Rhizoctonia spp. Isolated from Apple Roots and Orchard Soils

ABSTRACT Rhizoctonia spp. were isolated from the roots of apple trees and associated soil collected in orchards located near Moxee, Quincy, East Wenatchee, and Wenatchee, WA. The anastomosis groups (AGs) of Rhizoctonia spp. isolated from apple were determined by hyphal anastomosis with tester strains on 2% water agar and, where warranted, sequence analysis of the rDNA internal transcribed spacer region and restriction analysis of an amplified fragment from the 28S ribosomal RNA gene were used to corroborate these identifications. The dominant AG of R. solani isolated from the Moxee and East Wenatchee orchards were AG 5 and AG 6, respectively. Binucleate Rhizoctonia spp. were recovered from apple roots at three of four orchards surveyed and included isolates of AG-A, -G, -I, -J, and -Q. In artificial inoculations, isolates of R. solani AG 5 and AG 6 caused extensive root rot and death of 2- to 20-week-old apple transplants, providing evidence that isolates of R. solani AG 6 can be highly virulent and do not merely exist as saprophytes. The effect of binucleate Rhizoctonia spp. on growth of apple seedlings was isolate-dependent and ranged from growth enhancement to severe root rot. R. solani AG 5 and AG 6 were isolated from stunted trees, but not healthy trees, in an orchard near Moxee, WA, that exhibited severe symptoms of apple replant disease, suggesting that R. solani may have a role in this disease complex.     

Transformation of soil microbial community structure and rhizoctonia-suppressive potential in response to apple roots

ABSTRACT Changes in the composition of soil microbial communities and relative disease-suppressive ability of resident microflora in response to apple cultivation were assessed in orchard soils from a site possessing trees established for 1 to 5 years. The fungal community from roots of apple seedlings grown in noncultivated orchard soil was dominated by isolates from genera commonly considered saprophytic. Plant-pathogenic fungi in the genera Phytophthora, Pythium, and Rhizoctonia constituted an increasing proportion of the fungal community isolated from seedling roots with increasing orchard block age. Bacillus megaterium and Burkholderia cepacia dominated the bacterial communities recovered from noncultivated soil and the rhizosphere of apple seedlings grown in orchard soil, respectively. Populations of the two bacteria in their respective habitats declined dramatically with increasing orchard block age. Lesion nematode populations did not differ among soil and root samples from orchard blocks of different ages. Similar changes in microbial communities were observed in response to planting noncultivated orchard soil to five successive cycles of 'Gala' apple seedlings. Pasteurization of soil had no effect on apple growth in noncultivated soil but significantly enhanced apple growth in third-year orchard block soil. Seedlings grown in pasteurized soil from the third-year orchard block were equal in size to those grown in noncultivated soil, demonstrating that suppression of plant growth resulted from changes in the composition of the soil microbial community. Rhizoctonia solani anastomosis group 5 (AG 5) had no effect on growth of apple trees in noncultivated soil but significantly reduced the growth of apple trees in soil from third-year orchard soil. Changes in the ability of the resident soil microflora to suppress R. solani AG 5 were associated with reductions in the relative populations of Burkholderia cepacia and Pseudomonas putida in the rhizosphere of apple.     

复合微生物菌肥KMM减缓苹果再植病害机制

为揭示木美土里复合微生物菌肥（Kimidori microbial manure,KMM）促进再植苹果树的生长及减缓苹果再植病的作用机制,采用土壤常规农业化学分析法和Illumma MiSeq高通量测序技术比较分析KMM和常规有机肥（对照）处理后再植苹果树的生长指标、理化因子、根际土壤酶活性以及细菌群落结构和功能的变化。结果表明,施用KMM后,果树的株高增长率、干径增长率、叶绿素含量、分枝个数和分枝长度等生长指标均显著升高,有机质含量显著增加,中性磷酸酶、脲酶、蔗糖酶和过氧化氢酶活性均显著提高,同时增加了再植土壤细菌群落的丰富度和多样性。从采集土壤中分离得到51株细菌并进行盆栽试验,其中经KMM-15、KMM-37和KMM-50菌株处理后海棠苗叶绿素含量、株高、茎粗均显著提高,经鉴定这3株菌株为贝莱斯芽胞杆菌Bacillus velezensis。表明春秋2次施用KMM提高了土壤有机质含量和酶活性,并重塑了根际土壤中细菌群落结构,进而缓解苹果再植病害发生程度,促进果树生长。     

荧光假单胞菌SS101对苹果再植病害的生防效果

为明确荧光假单胞菌SS101对苹果再植病害的生防效果,采用平板对峙法和孢子萌发法研究了SS101菌株对9株来自不同苹果产区病原菌的抑制作用以及对盆栽海棠实生苗、海棠大苗和苹果树生长的影响,并测定了连续处理4年苹果树的产量。结果显示,SS101菌株对9株病原菌的菌丝生长抑制率为39.39%~72.00%,对孢子萌发抑制率为19.86%~51.77%;经SS101菌株处理后,海棠实生苗叶绿素含量、株高和鲜重均显著高于再植土处理,分别提高13.38%、19.39%和48.26%;经SS101菌株处理后,海棠大苗定植9个月后,茎粗净增长量为2.97 mm,增长率为214.05%,而再植土处理仅为114.02%;连续处理4年的苹果树平均单株产量比再植土处理提高139.46%,且消除了再植病害症状。研究表明,SS101菌株对苹果再植病害病原菌具有显著抑制作用,在田间对再植病害具有良好的防控效果。     

<Emphasis Type="Italic">Cylindrocarpon</Emphasis> species associated with apple tree roots in South Africa and their quantification using real-time PCR

Cylindrocarpon species are known to be a component of the pathogen/pest complex that incites apple replant disease. In South Africa, no information is available on apple associated Cylindrocarpon species and their pathogenicity. Therefore, these aspects were investigated. Among the isolates recovered from apple roots in South Africa, four species (C. destructans, C. liriodendri, C. macrodidymum and C. pauciseptatum) were identified using β-tubulin gene sequencing and phylogenetic analysis. This is the first report of C. liriodendri, C. macrodidymum and C. pauciseptatum on apple trees. Cylindrocarpon macrodidymum was the most prevalent. Isolates within each of the four species were pathogenic towards apple seedlings, but varied in their virulence. With a single exception, all isolates were able to induce lesion development on seedling roots. Only 57% of the isolates, which represented all four species, were able to cause a significant reduction in seedling weight and/or height. The greatest seedling growth reductions were caused by two isolates of C. destructans, and one isolate each of C. liriodendri and C. macrodidymum. A quantitative real-time polymerase chain reaction (qPCR) method was developed for simultaneous detection of all four Cylindrocarpon species. qPCR analyses of Cylindrocarpon from the roots of inoculated seedlings showed that the amount of Cylindrocarpon DNA in roots was not correlated to seedling growth reductions (weight and height) or root rot. The qPCR method is, however, very useful for the rapid identification of apple associated Cylindrocarpon species in roots. The technique may also hold potential for being indicative of Cylindrocarpon disease potential if rhizosphere soil rather than roots are used.