红麻种质资源根结线虫病抗性鉴定

为鉴定我国红麻种质资源对根结线虫病的抗病性,在对120份红麻遗传资源进行田间自然发病鉴定的基础上,选择25份有代表性(22份高抗和3份感病)的种质进行盆栽根结线虫病接种鉴定。田间自然发病鉴定结果:120份红麻遗传资源中5个高感、19个中感、25个中抗、71个高抗;其侵染的根结线虫主要为南方根结线虫(Meloidogyne incognita)、爪哇根结线虫(Meloidogyne javanica)和花生根结线虫(Meloidogyne arenria)的混合种群。进行盆栽接种鉴定的25份遗传资源中,7份为中抗,其余分别为中感和高感品种,未发现高抗或免疫品种。本研究可为红麻种质资源发掘利用提供科学依据。     

丛枝菌根真菌对西瓜嫁接苗抗南方根结线虫病的影响

根结线虫病是西瓜主要土传病害之一,长期连作往往加重西瓜根结线虫病害。利用抗病砧木嫁接可有效减轻瓜类蔬菜线虫病害,而接种丛枝菌根真菌(arbuscular mycorrhiza fungi,AMF)则能拮抗根结线虫,提高寄主植物抗病性。本试验旨在研究嫁接技术配合接种AMF降低瓜类蔬菜病害和促进生长的效应。试验以不同抗性南瓜杂交种(Cucurbita maxima×C.moschata)"青农2号"和"青农3号"分别嫁接西瓜Citrullus lanatus品种"京欣2号","青农2号"嫁接苗和"青农3号"嫁接苗先分别接种摩西管柄囊霉Funneliformis mosseae、变形球囊霉Glomus versiforme、根内根孢囊霉Rhizophagus intraradices,后接种南方根结线虫Meloidogyne incognita,测定AMF对不同西瓜嫁接苗根结线虫病和生长的影响。结果表明,接种AMF能不同程度地减轻嫁接苗根结线虫病害,其中,以"青农2号"嫁接苗接种变形球囊霉促生防病的效应最大。这可能与AMF提高根系活力、可溶性糖含量、超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性和过氧化氢酶(CAT)活性,降低丙二醛(MDA)含量有关。结果表明,"青农2号"嫁接苗结合接种变形球囊霉是生产上具有应用潜力的技术组合。     

葫芦科蔬菜种质资源对南方根结线虫的抗性评价

根据不同种质的来源地、农艺性状等背景信息,从国家蔬菜种质资源中期库中选取具代表性的444份主要瓜类作物地方品种,分属葫芦科7个属的13个种或变种,采用病土接种法进行苗期根结线虫抗性鉴定,得出了不同葫芦科作物对南方根结线虫的抗性分布范围。忽略基因型差异,不同作物的平均病级指数从小到大的顺序为:冬瓜、西瓜、丝瓜、节瓜、苦瓜、越瓜、甜瓜、菜瓜、瓠瓜、黄瓜、中国南瓜、印度南瓜、美洲南瓜。通过抗性鉴定,共获得27份抗根结线虫种质(病级指数1~2),包括12份冬瓜、3份苦瓜、7份丝瓜和5份西瓜。     

朱砂莲果实化学成分杀南方根结线虫活性研究

为了解朱砂莲(Aristolochia tuberosa O.F.Liang et S.M.Huang)的化学成分,从其果实的甲醇提取物中分离得到4个化合物。通过波普数据分析,分别鉴定为马兜铃内酰胺W(1)、8-epidiosbulbin e acetate(2)、diosbulbin B(3)和β-sitosterol(4)。化合物1~3对南方根结线虫2龄幼虫具有不同程度的毒杀作用,尤其马兜铃内酰胺W(1)活性最好,其96 h后的LC50为119.94μg m L–1。马兜铃属植物具有开发为新型植物源杀根结线虫剂的潜在价值。     

蓖麻提取物对南方根结线虫的防治作用

通过毒力测定及盆栽试验,研究了蓖麻提取物对南方根结线虫的杀线活性及防治效果.结果表明:蓖麻碱及蓖麻水提液均具有较强毒杀线虫活性,蓖麻碱浓度为2g·L-1、处理48 h杀线虫活性最强,线虫校正死亡率达91.5％,LC50为0.6 g·L-1;蓖麻水提液浓度为100g·L-1、处理48 h杀线虫活性最强,线虫校正死亡率达83.5％,LC50为18.3 g·L-1;蓖麻碱、蓖麻水提液和蓖麻叶植物粉处理接种南方根结线虫的番茄苗后,植株平均根结数分别为(17.6±1.7)、(20.6±1.5)和(22.8±3.7),均显著低于对照(37.4±2.3),根长分别比对照提高46.8％、34.5％和33.8％,株高分别比对照提高33.5％、22.6％和15.8％,植株鲜质量分别比对照增加41.4％、18.9％和10.1％.蓖麻提取物能减轻线虫危害,对盆栽番茄南方根结线虫病控制效果明显.     

13种植物源化合物对南方根结线虫的毒力比较

利用离体生物测定法比较了13种植物源化合物对南方根结线虫(Meloidogyne incognita Chitwood)的毒力,并探讨了紫外光照射对这些化合物杀虫活性的影响。结果表明,对苯二酚、DL-薄荷醇、丁子香酚和苦豆碱对南方根结线虫的毒杀活性最强,邻苯二酚等6种化合物的毒杀活性次之,野靛碱、毒扁豆碱和间苯二酚的毒力较低。紫外光照射可明显降低苦豆碱和柠檬酸对南方根结线虫的毒杀活性,而其他11种化合物的毒杀活性则基本不受紫外光照射的影响。     

南方根结线虫效应因子功能的研究进展

南方根结线虫(Meloidogyne incognita)是适生区分布广、为害普遍的一类植物内寄生线虫。线虫不同寄生阶段分泌的效应因子,在"植物-线虫"互作中,除了引起线虫的无毒特征外,也通过干扰植物正常的防御反应和细胞功能,进而参与线虫的早期侵染、迁移以及后期稳定寄生等过程。现从植物细胞壁的修饰、线虫无毒蛋白、植物免疫反应的抑制、生长素和细胞分裂素信号的调控,以及植物细胞结构和基础代谢的影响等方面,介绍南方根结线虫效应因子功能的研究现状。与此同时,对南方根结线虫效应因子功能研究中所存在的问题进行讨论,并展望未来的研究方向。     

南方根结线虫初始接种密度对生姜生长的影响

在盆栽条件下,研究了南方根结线虫不同初始接种密度对生姜生长的影响.结果表明,南方根结线虫的侵染,降低生姜株高、茎粗、分枝数、茎叶鲜重和根系鲜重,最终降低生姜的产量,且随着接种密度的增加,生姜生长所受到的危害也随之增加.每100 g干土接种0、100和200个卵,可使生姜分别减产27.91%、37.73%和42.14%.初始接种密度对南方根结线虫繁殖速率也有很大影响.一般初始接种密度低时繁殖速率高,接种密度高时繁殖速率低,其在生姜上的平衡密度为每100 g干土746.20个.     

丛枝菌根真菌对不同番茄品种抗根结线虫病的影响

【背景】根结线虫病害严重制约我国设施蔬菜的生产。丛枝菌根真菌(Arbuscular Mycorrhizal Fungi,AMF)作为土壤中最重要的有益真菌之一,可以促进植物生长,提高植物抗病性,减轻土传真菌和线虫病害的发生。在蔬菜保护地栽培中,AMF对于植物线虫病防治作用的研究受到了广泛关注。【目的】针对番茄生产中危害最严重的南方根结线虫(Meliodogyne incognita)病害问题,研究AMF和番茄品种不同组合的抗线虫效应,以期为菌根真菌作为生物防治剂和生物菌肥应用于实际生产提供技术基础。【方法】在灭菌土壤中,人工接种根结线虫,比较不同菌种Rhizophagus intraradices(Ri)、Acaulosporamellea(Am)及菌种组合Rhizophagusintraradices+Acaulosporamellea(Ri+Am)在不同番茄品种(感病品种蒙特卡罗和抗线虫品种仙客1号)上对南方根结线虫侵染和繁殖的影响,研究AMF对根结线虫的拮抗效应。另外,采用南方根结线虫连作发病的土壤,在感病品种蒙特卡洛上接种AMF混合菌种Ri+Am,番茄苗移栽入连作土壤中,测定各生长指标和调查根结和卵块数量,评价接种AMF处理对根结线虫病的防治效果。【结果】在灭菌土壤中,普通番茄品种蒙特卡罗的菌根效应显著优于抗线虫番茄品种仙客1号,表现为前者单位根重的根结和卵块的数量均比对照显著降低,而后者仅降低了卵块数量;蒙特卡罗上接种Ri+Am混合菌种的效果优于接种单一菌种Am和Ri;而仙客1号上接种Ri的效果更好。接种线虫也显著影响了AMF的侵染,而且对抗性品种仙客1号的影响更为明显。但除了接种Am的处理,大多数处理收获时菌根侵染率仍维持较高的水平(70%以上)。在连作土壤中,感病品种蒙特卡罗接种混合菌种Ri+Am具有较好的抗/耐线虫效应,主要表现为促进植株生长,显著降低根结和卵块数量,但菌根侵染率较灭菌土壤低,约为40%。【结论】综合以上结果,表明菌根化苗能够在一定程度上减轻根结线虫病的危害。土壤灭菌条件下,在感病和抗线虫番茄品种上接种AMF能够减轻线虫的侵染和繁殖,而且在感病品种上的防治效果更加显著。在连作土壤中,在番茄感病品种上接种AMF也表现较好的抗线虫效果。     

基于GIS的南方根结线虫在陕西省越冬区划分析

土壤温度是影响南方根结线虫(Meloidogyne incognita)越冬的重要因子。通过自动温度记录仪,从2009—2012年的每年冬季(11月至次年3月),对陕西省延安、商洛、杨凌和大荔4个生态区的气温和不同种植模式下的土壤温度进行数据采集和模拟统计,建立土温与气温关系的数学模型,根据模型将陕西省96个气象站点的气温数据转换为土温数据。利用GIS的克里金(Kriging)空间插值功能,结合实验室测得的南方根结线虫存活的最低温度,生成南方根结线虫在拱棚、地膜覆盖和露地3种种植条件下的越冬区划图并进行分析。研究表明,(1)0℃以下低温对南方根结线虫有明显抑制作用,南方根结线虫在低于-1℃低温持续32 d以上时无法越冬。(2)土壤温度和气温呈线性相关关系,4种种植条件下土壤温度(Y)与气温(X)的关系方程分别为露地:Y=0.8125X+1.9325,R=0.934;地膜覆盖:Y=0.7943X+1.8563,R=0.918;拱棚:Y=0.7046X+6.2685,R=0.907;温室:Y=0.302X+14.519,R=0.597。(3)最冷月土壤均温低于-1℃的概率在70%—80%的区域可以认为是南方根结线虫的越冬界线,越冬界线在露地、地膜覆盖和拱棚条件下依次北移,在温室条件下可在全省范围内越冬。     

Hatching Response of Meloidogyne incognita acrita to Electric Shock

The influence of electric shock on hatch of Meloidogyne incognita acrita from egg masses taken from roots of ''Acala SJ-I'' cotton (Gossypium hirsutum L.) was studied. Egg masses in tap water were individually placed between the tips of needle electrodes 1 mm apart and exposed to potentials of l, 10, 20, and 60 vdc/mm at 1, 1, 1, and 86 milliamperes dc, respectively, for periods of 2 and 60 seconds. Hatched larvae were counted at five-day intervals for 60 days. Of the eight treatment combinations used, six gave a greater egg hatch than the control. The largest hatch, 520 percent greater than the control, resulted from exposure to 1 vdc/mm for 60 seconds; 60 vdc/mm for 2 and 60 sec decreased egg hatch 11 and 94 percent of the untreated control. Hatched larvae from all treatments except the 60 vdc/mm, 60-second exposure were infective and reproduced on young cotton plants in a glasshouse.     

Differential expression of root-knot nematode resistance genes in tomato and pepper: evidence with Meloidogyne incognita virulent and avirulent near-isogenic lineages

Reproduction of artificially selected near isogenic Meloidogyne incognita lineages virulent and avirulent against the Mi resistance gene of tomato was assessed on host and resistant lines and cultivars of pepper. Egg mass production following inoculation of individual potted seedlings with second-stage juveniles was studied in experiments conducted in controlled environment. Artificially selected Mi-virulent nematode populations were unable to develop on resistant pepper lines PM 217 and PM 687. This suggests that the genetic systems governing resistance to root-knot nematodes are differently expressed in tomato and pepper, in spite of the very close phylogenetic relationships and structural genomic homologies occurring between these two vegetable crops. Moreover, these artificially selected nematode populations were also found unable to develop on the susceptible pepper cultivars California Wonder and Doux Long des Landes, while their pathogenicity was not significantly affected on susceptible tomatoes. Due to the existence of naturally virulent Meloidogyne populations, these results enhance the need for a better understanding of the mechanisms involved, in order to develop new forms of management of plant resistance to root-knot nematodes.     

System optimization for attachment of Pasteuria penetrans to Meloidogyne incognita

For optimal mass production of Pasteuria penetrans in vivo, it is important to develop a system that can ensure 100% nematode attachment of the bacteria and high bacterial infection after inoculation. In this study, effects of endospore concentration and centrifugation parameters on attachment were investigated, followed by evaluation of impacts of centrifugation on endospore dislodgement, Meloidogyne incognita juvenile (J2) mortality, J2 infectivity, and bacterial infectivity. Endospore concentration and percentage of attachment fit well to mass-action and logit models, with the former being superior. Centrifugation had no impact on J2 mortality but had a great impact on endospore dislodgement in sand and water, nematode infectivity and bacterial infectivity. At nematode concentration of 2×10³ J2/mL, the optimal system for endospore attachment was developed which consisted of bacteria at 2×10⁴ endospores/mL, and centrifugation at 9000×g for 3 min three times. This system generated 100% attachment with approximately seven endospores/J2. After inoculation of treated nematodes to tomato plants, the inoculum yielded 47% bacterial infection, superior to 17% infection observed in centrifugation at 6000×g. Endospore dislodgement occurred after placing the centrifuged inoculated nematodes in sand or water for 24 and 48 h, which was more severe in centrifugation at 6000 than at 9000×g. Results also indicated that centrifugation led to lower nematode infectivity, regardless of endospore presence and centrifugation at 9000 or 6000×g, compared with the no centrifugation control.     

In vitro studies of antagonistic fungi against the root-knot nematode,Meloidogyne incognita

The present study was carried out in vitro to determine the efficacy of indigenous fungi isolated from egg masses of root-knot nematode, Meloidogyne incognita on egg parasitism, egg hatching, mobility and mortality against root-knot nematode, M. incognita. The tested fungi were Acremonium strictum, Aspergillus terreus, A. nidulans, A. niger, Chetomium aubense, Chladosporium oxysporum, Fusarium chlamydosporium, F. dimarum, F. oxysporum, F. solani, Paecilomyces lilacinus, Pochonia chlamydosporia, Trichoderma viride and T. harzianum. All tested fungi showed varied effects against the nematodes. Culture filtrates of A. strictum was very effective against the nematode in regards to egg parasitism (53%), egg hatching inhibition (86%) and mortality (68%) compared to controls. A. strictum was found to have an advantage over P. lilacinus, P. chlamydosporia, T. viride and T. harzianum in that it caused greater mortality of the second stage juveniles (J₂). A. terreus did not show egg parasitism but was found to be highly toxic against second stage juveniles (J₂) causing high mortality (around 68%). Thus, A. strictum and A. terreus showed good biocontrol potential against root-knot nematode, M. incognita under in vitro conditions.     

Histopathology of Root-knot Nematode (Meloidogyne incognita) Infection on White Yam (Dioscorea rotundata) Tubers

White yam tissues naturally and artificially infected with root-knot nematodes were fixed, sectioned, and examined with a microscope. Infective second-stage juveniles of Meloidogyne incognita penetrated and moved intercellularly within the tuber. Feeding sites were always in the ground tissue layer where the vascular tissues are distributed in the tubers. Giant cells were always associated with xylem tissue. They were thin walled with dense cytoplasm and multinucleated. The nuclei of the giant cells were only half the size of those found in roots of infected tomato plants. Normal nematode growth and development followed giant cell formation. Females deposited eggs into a gelatinous egg mass within the tuber, and a necrotic ring formed around the female after eggs had been produced. Second-stage juveniles hatched, migrated, and re-infected other areas of the tuber. No males were observed from the tuber.     

Nicotine Content of Tobacco Roots and Toxicity to Meloidogyne incognita

The motility of Meloidogyne incognita second-stage juveniles (J2) and their ability to induce root galls in tomato were progressively decreased upon exposure to nicotine at concentrations of 1-100 μg/ml. EC₅₀ values ranged from 14.5 to 22.3 μg/ml, but J2 motility and root-gall induction were not eliminated at 100 μg/ml nicotine. Nicotine in both resistant NC 89 and susceptible NC 2326 tobacco roots was increased significantly 4 days after exposure to M. incognita. The increase was greater in resistant than in susceptible tobacco. Root nicotine concentrations were estimated to be 661.1-979.1 μg/g fresh weight. More M. incognita were detected in roots of susceptible than in roots of resistant tobacco. Numbers of nematodes within resistant roots decreased as duration of exposure to M. incognita was increased from 4 to 16 days. Concentrations of nicotine were apparently sufficient to affect M. incognita in both susceptible and resistant tobacco roots. Localization of nicotine at infection sites must be determined to ascertain its association with resistance.     

The role of chitinase from Lecanicillium antillanum B-3 in parasitism to root-knot nematode Meloidogyne incognita eggs

B-3 fungal isolate was isolated from soil samples of Gwangju in Korea. Based on morphological and phylogenetic analysis, it was designated as Lecanicillium antillanum B-3 (syn. Verticillium antillanum B-3). The fungus was a chitinolytic-nematophagous microorganism. B-3 chitinase activity from 0.5% swollen chitin broth medium reached the highest level on the sixth day and then plateaued until 12 days. B-3 isolate showed the high rate of parasitism on Meloidogyne incognita eggs with more than 90% infection rate on the third day after treatment. B-3 crude chitinase damaged the eggshell structures more than 78% based on lactoglycerol staining data at a final protein concentration of 14.6 µg mL^?1 on the fourth day following treatment. Partially purified chitinase with molecular 37 kDa from DEAE-Sephadex chromatography also showed damaging effect on the eggs. These results suggested that chitinase from B-3 isolate was responsible for degradation of M. incognita eggshell structures.     

Extremely Sensitive Thermotaxis of the Nematode Meloidogyne incognita

Eggs of the root-knot nematode Meloidogyne incognita were acclimated to 23 C. Newly hatched second-stage juveniles migrated toward higher temperatures when placed in shallow thermal gradients averaging 23 C. The threshold gradient for this response was below 0.001 C/cm, with a best estimate of 4 x 10⁻⁴ C/cm. Calculations of physical limitations on thermotaxis indicate that this sensitivity is well within the limits of what is physically possible.