土壤强还原消毒过程中产生气体对土传病原菌的抑制作用

土壤强还原(reductive soil disinfestation,RSD)是一种高效、环保的土壤消毒方法,为明确RSD杀菌机理,采用气相色谱和荧光定量PCR等技术研究RSD过程中产生的气体对土传病原菌的杀菌作用。结果表明:RSD过程中土壤能够产生H_2S和NH_3;当在装有25 g病土的三角瓶中分别加入25 m L的H_2S和NH_3时,土壤中尖孢镰刀菌的数量均显著下降,其杀菌效果分别为90.3%和100.0%;当在病土中加入0.125%(w/w)氨水时,尖孢镰刀菌、茄劳尔氏菌、辣椒疫霉和立枯丝核菌的数量分别下降至对照土壤的1.0%、0.3%、0.1%和0.9%;此外,加入10μL氨水即可显著抑制尖孢镰刀菌菌丝生长和孢子萌发,抑制率分别为52.8%和100.0%。表明RSD过程中产生的H_2S和NH_3对RSD过程的杀菌效果起着一定的作用。     

土壤施加氧化钙对西瓜枯萎病的影响及其机理初探

为探明土壤撒施石灰对西瓜枯萎病发生的影响,本试验设计5个浓度剂量的氧化钙对连作西瓜5年的土壤进行消毒处理,通过栽培感病品种并调查植株生长情况、枯萎病发病率等指标,结合土壤中尖孢镰刀菌西瓜专化型1号小种病原菌孢子量和土壤pH的变化动态,以此评估土壤施加石灰防治西瓜枯萎病的效果,并利用不同浓度的钙对尖孢镰刀菌西瓜专化型菌丝生长和产孢量的影响进行防治机理的初步研究。试验结果表明,当土壤施加氧化钙后,西瓜发病率显著降低,各处理的相对防效分别为14%、57.8%、96.3%、94.7%、94.7%。在定植10d后,各处理均有促进植株生长发育的作用,其中1.34g/kg最显著。而随着氧化钙施加量的增加,土壤pH值也不断提高,30d趋于稳定。对土壤病原菌孢子数量检测表明:各处理土壤中病原菌数量均呈现显著降低的趋势。PDA平板生测结果表明,当钙离子浓度高于80mmol/L时,病原菌的生长及产孢量开始受到抑制,随着浓度的增加,抑制作用加强。土壤施加氧化钙不仅能改善土壤的酸化,还能有效防控西瓜枯萎病,其机理可能是土壤pH值的提高改变了土壤微环境,不利于病原菌侵染;施加的钙离子抑制了病原菌菌丝的生长及孢子的萌发。研究结果对揭示氧化钙防控西瓜枯萎病机制,具有一定的参考价值。     

烟草尖孢镰刀菌拮抗真菌的筛选鉴定及促生作用研究

为获得烟草镰刀菌根腐病拮抗真菌菌株,试验以河南省豫中烟区烟草根际土壤为试材,以主要致病菌尖孢镰刀菌Fusarium oxysporum为主要靶标,采用稀释涂布平板法分离获得土壤真菌371株,平板对峙培养法测定获得1株具有明显抑制效果的拮抗菌株,利用形态学和分子生物学的方法将其鉴定为棘孢木霉Trichoderma asperellum Tr-0111。抑制效果测定表明,该菌株对尖孢镰刀菌的抑制率最高可达93.13%,对根串珠霉菌、拟茎点霉、链格孢菌等其他8种烟草病原菌均具有较强的抑制效果。促生作用测定结果表明,菌株发酵液使烟草种子萌发率提高5.34%,烟苗根长增长率为62.39%,对盆栽烟草的最大叶面积和地上部鲜重也具有显著的促生效果。为进一步相关机理的研究和生防菌剂的研制奠定了基础。     

大白菜枯萎病病原镰刀菌种类的初步研究

 2014~2016年,由于栽培管理及重茬等原因,大白菜枯萎病在我国大面积发生,造成了巨大的经济损失。为了明确引起大白菜枯萎病的病原菌,本课题组从山东、内蒙古、河北、甘肃等大白菜主产区采集了具有典型枯萎病症状的病样,并对样品中的病原菌进行了分离和鉴定。形态学鉴定结果表明:分离物分别具有尖孢镰刀菌 (Fusarium oxysporum)、茄病镰刀菌 (F. solani) 和木贼镰刀菌 (F. equiseti)的形态学特征。柯赫氏法则验证结果表明:3种病原菌均能使大白菜发病,且发病症状与田间症状一致。此外,基于病原菌的rDNA-ITS和mt SSU序列的测序比对,3种病原菌与尖孢镰刀菌、茄病镰刀菌和木贼镰刀菌的同源性分别达99%~100%,这与形态学鉴定结果相一致。尖孢镰刀菌引起白菜枯萎病为国内首次报道,而茄病镰刀菌和木贼镰刀菌引起白菜枯萎病为国内外首次报道。     

藏红花球茎腐烂病病原鉴定及防治药剂室内筛选

球茎腐烂病是严重影响藏红花球茎品质和柱头产量的病害。为鉴定引起藏红花球茎腐烂病的病原菌及筛选防治该病害的有效杀菌剂,本研究基于形态学特征、rDNA-ITS和TEF-1α序列分析,对青海省藏红花球茎腐烂病的病原菌进行了鉴定,并利用菌丝生长速率法测定了10种杀菌剂对病原菌的抑菌作用。结果发现,引起藏红花球茎腐烂病的病原菌为尖孢镰刀菌Fusarium oxysporum和木贼镰刀菌F. equiseti,其中尖孢镰刀菌为优势病原菌。室内药剂试验表明70%甲基硫菌灵WP、 50%咯菌腈WP、 50%苯醚甲环唑SC和25%硅唑·咪鲜胺EW对两种镰刀菌均具有较好的抑制作用,EC50在0.574 0~1.808 6 mg/L之间,64%噁霜·锰锌WP、 30%噁霉灵AS和45%石硫合剂WP对2种镰刀菌的抑菌作用较差,EC50在2.134 4~7.915 3 mg/L之间。试验结果为生产上合理选用杀菌剂防治藏红花球茎腐烂病提供了科学依据。     

甘肃定西地区甘蓝枯萎病病原菌的分离与鉴定

自2009年起,甘肃定西地区出现了甘蓝植株矮化、叶片黄化、枯萎甚至死亡的现象。2015年8月,我们采集了田间病株样本,使用常规组织分离法对病原菌进行了分离和纯化,依据柯赫氏法则进行了病原菌确认,并通过形态学和分子生物学方法对病原菌进行了鉴定。结果表明病原菌的形态学特征与尖孢镰刀菌Fusarium oxysporum一致,其rDNA-ITS、rDNA-IGS以及EF-1α序列与尖孢镰刀菌F.oxysporum相似性达99%,基于病原菌及尖孢镰刀菌各代表专化型EF-1α序列构建的系统发育树将该菌与尖孢镰刀菌黏团专化型F.oxysporum f.sp.conglutinans聚为一类,故引致甘肃定西地区甘蓝枯萎病的病原菌为尖孢镰刀菌黏团专化型F.oxysporum f.sp.conglutinans。     

巴拿马进口香蕉上的尖孢镰刀菌对玉米苗期侵染试验简报

在巴拿马进口香蕉上分离获得的尖孢镰刀菌,编号为 NP1800,哈尔滨医科大学和中国科学院微生物所合作进行“串珠镰刀菌和尖孢镰刀菌对玉米苗期侵染的接种试验”,用该菌作试验菌种,在他们试验的9种尖孢镰刀菌中,NP1800是对玉米幼苗致病性最强的菌株。尖孢镰刀菌是广寄主专化型的。我们认为该菌除为害香蕉之外,还有可能为害其他农作物。因此我们进行了尖孢镰刀菌对玉米苗期的侵染试验。获得了以下4方面的数据:(1)不同玉米品种注射接种的发病情况(2)尖孢镰刀菌对不同玉米品种发病回收统计(3)注射接     

河北廊坊大豆枯萎病病原镰刀菌的分子鉴定

 为明确河北廊坊中国农科院植保所试验基地大豆孢囊线虫病田内大豆枯萎病病原镰刀菌的种类,对362份罹病枯萎大豆植株进行病原真菌分离,得到335株真菌;使用镰刀菌通用引物鉴定出镰刀菌(Fusarium spp.) 279株,占分离菌株83.3%;镰刀菌特异性引物、测序等分子生物学技术结合形态学特征进一步鉴定镰刀菌种类,鉴定出尖孢镰刀菌(F. oxysporum)189株,占分离菌株56.4%;茄病镰刀菌(F. solani)67株占20.0%、禾谷镰刀菌(F. graminearum)16株占4.8%、木贼镰刀菌(F. equiseti)3株、层出镰刀菌(F. proliferaum)2株、燕麦镰刀菌(F. avenaceum)和厚孢镰刀菌(F. chlamydosporum)各1株;致病性测试结果表明数量最多的尖孢镰刀菌(F. oxysporum)中约92.8%菌株具有不同程度的致病力;这些结果表明该试验基地大豆枯萎病的优势病原菌为尖孢镰刀菌(F. oxysporum);研究结果可为大豆枯萎病的防治提供科学依据,并为大豆孢囊线虫与尖孢镰刀菌复合侵染大豆的研究奠定基础。     

具防病并提高植物耐旱功能的小链霉菌HS57的筛选

本研究旨在寻找防治黄瓜枯萎病并提高植物抗逆能力的多功能菌株。从连作田健康黄瓜根际土中分离筛选到1株放线菌HS57。平板对峙法测定菌株HS57对尖孢镰刀菌、茄病镰刀菌和立枯丝核菌的抑制作用,平皿对扣共培养法测定其挥发性物质对尖孢镰刀菌的抑制作用,温室盆栽法测定其对黄瓜枯萎病的防病效果及对小麦耐旱能力的影响,结合菌株形态特征和16S rRNA基因序列分析对其进行鉴定。结果表明,菌株HS57可抑制3种病原菌生长,其中对尖孢镰刀菌的抑制率为52.4%;其挥发性物质对尖孢镰刀菌的抑制率为32.1%。浓度为10^7个/mL的HS57孢子悬浮液浸种对黄瓜枯萎病的防效最好,为51.0%;小麦种子经浓度为10^7、10^8个/mL的HS57孢子悬浮液浸种,麦苗经连续7 d缺水处理后再浇水,比对照恢复得更好。初步鉴定HS57菌株为小链霉菌Streptomyces parvus,是一株非常有潜力的多功能菌株。     

一株合欢枯萎病拮抗菌的分离鉴定及盆栽防治试验

合欢枯萎病是导致多年生合欢衰弱和死亡的主要因素之一,目前尚无有效的防治手段。本研究通过平板对峙法和含毒培养基法分离筛选健康合欢植株根际土壤中的潜在拮抗菌,结合形态学特征和双基因系统发育分析对拮抗菌进行鉴定,并通过盆栽试验验证拮抗菌株对合欢枯萎病的防治效果。结果表明,经过初筛得到11株对合欢枯萎病具有潜在拮抗作用的菌株,其中菌株5-2的抑制作用最为显著。对峙培养6d,菌株5-2对尖孢镰刀菌Fusariumoxysporum和腐皮镰刀菌Fusariumsolani的抑制率分别为76.88%和84.08%,两种病原菌在含6.67%拮抗菌孢悬液(孢子浓度1.0×10⁷个/mL)的平板上生长均受到显著抑制。显微观察发现菌株5-2可缠绕病原菌菌丝体并寄生生长,盆栽试验表明菌株5-2对尖孢镰刀菌和腐皮镰刀菌具有优良的拮抗作用,防治效果达78.26%和57.89%。通过形态学和ITS-TEF1双基因系统发育分析,菌株5-2鉴定为康宁木霉Trichodermakoningii。综上所述,菌株5-2对合欢枯萎病具有良好的防治效果,可作为合欢枯萎病生防菌剂的候选菌株资源。     

Role of organic acids in the mechanisms of biological soil disinfestation (BSD)

Biological soil disinfestation (BSD), or reductive soil disinfestation, achieved by amendment with organic materials such as wheat bran followed by flooding and covering the soil surface, has been used to control some soilborne diseases including Fusarium wilt and bacterial wilt of tomato. During a BSD treatment, accumulation of acetic acid and/or butyric acid was detected with high-performance liquid chromatography. Survival of Fusarium oxysporum f. sp. lycopersici or Ralstonia solanacearum was suppressed by these organic acids. Amendment of these organic acids into soil suppressed the survival of R. solanacearum at lower concentrations than the maximum detected in BSD treatment, indicating that production of these organic acids is one of the mechanisms of control. However, F. oxysporum f. sp. lycopersici in soil survived with the maximum concentrations of these organic acids achieved by BSD; thus, involvement of factors other than organic acids may be involved.     

尖镰孢菌(Fusarium oxysporum)的快速分子检测

 由尖镰孢菌(Fusarium oxysporum Schlecht.)引起的大豆枯萎病是危害大豆生产的主要土传病害^［1］。该菌在土壤和病残体上均可长期生存造成危害。快速准确地在发病初期植株和带病土壤中进行鉴定和检测对防治该病害至关重要。     

Survival of soil-borne fungus Phymatotrichopsis omnivora after exposure to volatile fatty acids

Control of plant-pathogenic Phymatotrichopsis omnivora is difficult. In tests of volatile fatty acids (VFAs), mycelia in empty 9-cm Petri dishes with 3 or 6 μL of acetic, butyric, formic or propionic acid did not grow, but 37–67 % grew on potato dextrose agar with 6 μL VFA. Survival of sclerotia was <20 % on water agar with 154 μg/mL of a VFA or 12–40 % in sandy soil with 616 μg/mL. Survivability after 2-h immersion in 17 mmol/L varied greatly (0–100 %) depending on the VFA, but in 33–50 mmol/L, 0 % survived. This fungus was thus susceptible to VFAs.     

低分子量有机酸对不同肥力土壤磷素的活化作用

针对石灰性土壤磷素固定累积问题,采用化学浸提方法,探讨低分子有机酸对不同肥力水平土壤磷素的活化作用,提高磷素的有效性.结果表明,对于低肥力和中肥力土壤,相同浓度下有机酸活化土壤磷的能力表现为草酸＞柠檬酸＞苹果酸;对于高肥力土壤,相同浓度下有机酸活化土壤磷的能力为柠檬酸＞草酸＞苹果酸.低浓度对有机骏对土壤磷的活化有抑制作...     

Corn straw return can increase labile soil organic carbon fractions and improve water-stable aggregates in Haplic Cambisol

Corn straw return to the field is a vital agronomic practice for increasing soil organic carbon (SOC) and its labile fractions, as well as soil aggregates and organic carbon (OC) associated with water-stable aggregates (WSA). Moreover, the labile SOC fractions play an important role in OC turnover and sequestration. The aims of this study were to determine how different corn straw returning modes affect the contents of labile SOC fractions and OC associated with WSA. Corn straw was returned in the following depths: (1) on undisturbed soil surface (NTS), (2) in the 0-10 cm soil depth (MTS), (3) in the 0-20 cm soil depth (CTS), and (4) no corn straw applied (CK). After five years (2014-2018), soil was sampled in the 0-20 and 20-40 cm depths to measure the water-extractable organic C (WEOC), permanganate oxidizable C (KMnO₄-C), light fraction organic C (LFOC), and WSA fractions. The results showed that compared with CK, corn straw amended soils (NTS, MTS and CTS) increased SOC content by 11.55%-16.58%, WEOC by 41.38%-51.42%, KMnO₄-C and LFOC by 29.84%-34.09% and 56.68%-65.36% in the 0-40 cm soil depth. The LFOC and KMnO₄-C were proved to be the most sensitive fractions to different corn straw returning modes. Compared with CK, soils amended with corn straw increased mean weight diameter by 24.24%-40.48% in the 0-20 cm soil depth. The NTS and MTS preserved more than 60.00% of OC in macro-aggregates compared with CK. No significant difference was found in corn yield across all corn straw returning modes throughout the study period, indicating that adoption of NTS and MTS would increase SOC content and improve soil structure, and would not decline crop production.     

低分子量有机酸对二氯喹啉酸在土壤中吸附-解吸的影响

采用高效液相色谱仪及批量平衡试验方法,研究了乙酸、苹果酸、酒石酸、草酸、丁二酸和柠檬酸6种低分子量有机酸对麻沙泥和第四纪红土红壤吸附-解吸二氯喹啉酸的影响。结果表明:低分子量有机酸可推迟二氯喹啉酸在土壤中的吸附平衡时间,其吸附动力学过程可用准二级动力学方程描述。Linear和Freundlich方程能较好地拟合二氯喹啉酸在供试两种土壤中的吸附等温线;二氯喹啉酸在麻沙泥中的吸附能力(lg Kf值)从大到小依次为苹果酸柠檬酸草酸=乙酸丁二酸酒石酸,在第四纪红土红壤中为苹果酸丁二酸乙酸草酸柠檬酸酒石酸;低分子量有机酸浓度对二氯喹啉酸解吸的影响因有机酸种类和供试土壤的不同而差异较大,6种供试有机酸均促进了第四纪红土红壤对二氯喹啉酸的解吸,且其解吸率均明显高于麻沙泥对二氯喹啉酸的解吸率,但在麻沙泥中呈现不同的影响模式。     

Transgenically enhanced expression of indole-3-acetic Acid confers hypervirulence to plant pathogens

ABSTRACT Fusarium oxysporum and F. arthrosporioides, pathogenic on Orobanche aegyptiaca, were transformed with two genes of the indole-3-acetamide (IAM) pathway leading to indole-3-acetic acid (IAA) to attempt to enhance virulence. Transgenic F. oxysporum lines containing both the tryptophan-2-monooxyngenase (iaaM) and indole-3-acetamide hydrolase (iaaH) genes produced significantly more IAA than the wild type. IAM accumulated in culture extracts of F. oxysporum containing iaaM alone. F. arthrosporioides containing only iaaM accumulated IAM and an unidentified indole. Some transformants of F. oxysporum expressing only the iaaM gene also produced more IAA than the wild type. Sub-threshold levels (that barely infect Orobanche) of transgenic F. oxysporum expressing both genes and of F. arthrosporioides expressing iaaM were more effective in suppressing the number and size of Orobanche shoots than the wild type on tomato plants grown in soil mixed with Orobanche seed. Stimulating an auxin imbalance enhanced pathogen virulence by affecting the host in a manner similar to low doses of auxin herbicides such as 2,4-dichlorophenoxy acetic acid.     

载银无机抗菌制剂对大豆根腐病的防治效果

 测定了广泛用于工业和日用品的载银无机抗菌制剂Zeomic和AM₁对大豆根腐病重要病原菌Fusarium oxysporum、Fusarium solani和Rhizoctonia solani孢子萌发和菌落扩展的抑制作用。结果表明,Zeomic在浓度为500 mg/L时可以完全抑制F. oxysporum和F. solani的孢子萌发,但对F. oxysporum、F. solani和R. solani菌落扩展的抑制作用较弱,分别为7.4%、0.0%和3.7%。AM₁对病菌孢子萌发的抑制作用随浓度的升高而增强,在浓度为250 mg/L时对F. solani孢子萌发的抑制率为18.8%,到1 000 mg/L时可完全抑制。但AM₁在浓度为1 000 mg/L时对F. oxysporum的孢子萌发抑制率只有29.7%,表明不同菌类对AM₁的敏感性不同。同一浓度下的AM₁对F. oxysporum菌落扩展的抑制率大于对F. solani和R. solani菌落扩展的抑制作用,在500 mg/L时对病菌的抑制率分别为51.4%、14.6%和11.0%。温室内对大豆生长的调查结果表明,用种子重量0.05%和0.10%的AM₁拌种对出苗和生长有促进作用,Zeomic用量在0.05%时不影响出苗、株高和根系干物质积累,但当用量增加至0.10%时,则表现对出苗和干物质累积的抑制作用。田间试验结果表明,500m g/L的AM₁拌种对大豆根腐病防效为32.6%,与化学农药35%多克福包衣处理无显著差异,但比后者增产10.1%。由此说明,载银无机抗菌制剂具有用于农作物病害防治的潜力。     

土壤拮抗放线菌s-930-6菌株活性产物抑菌作用

研究结果表明,用生长速率法测定自秦岭太白山区土壤中分离出的拮抗放线菌s-930-6菌株,其发酵液粗提物对供试病原菌都有不同程度的抑制作用,其中对小麦根腐病菌的菌丝生长作用最强,EC50为2.6g/L.用孢子萌发法测定,该粗提物对黄瓜枯萎病菌和番茄枯萎病菌的孢子萌发抑制作用较强,EC50分别为0.04、0.02g/L.用D101大孔树脂对粗提物进行提取得到精提物,它对5种供试病菌的菌丝生长在500mg/L浓度下有不同程度的抑制作用,其中对小麦赤霉病菌、小麦根腐病菌和黄瓜枯萎病菌的抑制效果超过85%.精提物在500mg/L浓度下对小麦白粉病的保护作用为76.39%,对黄瓜霜霉病保护作用为82.38%,治疗作用为78.31%.用高效液相色谱(HPLC)分离纯化精提物得到4个组分,其中B组分对小麦赤霉病菌的抑制作用最强,达到94.14%,初步鉴定B组分为氨基糖苷类化合物.