Systemically induced resistance and microbial competitive exclusion: implications on biological control

The root-knot nematode, Meloidogyne incognita, is among the most damaging agricultural pests, particularly to tomato. The mutualistic endophytes Fusarium oxysporum strain Fo162 (Fo162) and Rhizobium etli strain G12 (G12) have been shown to systemically induce resistance toward M. incognita. By using triple-split-root tomato plants, spatially separated but simultaneous inoculation of both endophytes did not lead to additive reductions in M. incognita infection. More importantly, spatially separated inoculation of Fo162 and G12 led to a reduction in Fo162 root colonization of 35 and 39% when G12 was inoculated on a separate root section of the same plant in two independent experiments. In an additional split-root experiment, spatial separation of Fo162 and G12 resulted in a reduction of Fo162 root colonization of approximately 50% over the water controls in two independent experiments. The results suggested that the suppressive activity of G12 on Fo162 and M. incognita is possibly related to the induction of specific plant defense mechanisms. Thus, although Fo162 and G12 have the ability to systemically repress M. incognita infection in tomato, they can be considered incompatible biocontrol agents when both organisms are present simultaneously on the same root system.     

模拟酸雨对小麦条锈病流行学组分的影响

为明确酸雨对小麦条锈病菌孢子萌发及病害流行学组分的影响,采用不同酸度的模拟酸雨(p H值分别为2.5、3.5、4.5和5.6)处理小麦条锈病菌Puccinia striiformis f.sp.tritici生理小种CYR32和CYR33的夏孢子,并将不同酸度模拟酸雨处理后的夏孢子接种小麦感病品种铭贤169,采用组分分析法研究酸雨对病害发展过程的影响。结果表明,随着模拟酸雨酸度的增强,生理小种CYR32和CYR33的夏孢子萌发率均显著降低,萌发24 h后对照组生理小种CYR32和CYR33的夏孢子萌发率分别为93.7%和79.8%,在p H 3.5模拟酸雨处理后萌发率仅为13.1%和8.6%。温室内接种试验结果显示,不同酸度模拟酸雨对生理小种CYR32和CYR33的侵染概率、潜育期和病情指数均有显著影响,对生理小种CYR32和CYR33病斑扩展率影响不显著;与对照相比,p H 3.5模拟酸雨处理后生理小种CYR32和CYR33的侵染概率分别下降79.9%和79.8%。结果表明强酸雨(p H 3.5)显著延长小麦条锈菌潜育期,减少病斑产孢量,降低病情指数和病害进展曲线下面积。     

AM真菌诱导的番茄信号物质及其对根结线虫的 抑制 效应

丛枝菌根(AM)真菌能诱导植物合成一些信号物质,如茉莉酸(JA)、水杨酸(SA)、一氧化氮(NO)、H2O2等,这些信号在AM真菌与植物识别、共生体建立和激活植物防御系统过程中发挥着重要作用[1].     

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。     

Management of root-knot disease of tomato by the application of fly ash in soil

The effects of fly ash at different concentrations (0, 10, 20, 30 … 100% vol./vol. in soil) on plant growth and yield were investigated in tomato plants infected or noninfected with root-knot nematode, Meloidogyne incognita (2000 juveniles per plant) in clay pots. An increase in fly ash concentration in the soil correspondingly increased the availability of carbonates, bicarbonates, sulphate, chlorides, B, P, K, Ca, Mg, Mn, Cu and Zn in the soil. The porosity, water-holding capacity, pH, conductivity and cation exchange capacity also increased progressively in the fly ash amended soil. Ash application enhanced plant growth, leaf pigment concentrations, fruit production, weight of fruit/plant and mean fruit weight of both nematode-infected and noninfected tomato plants, being maximum in the soil containing 50 or 60% fly ash. The yield enhancements were 93.6 (infected plants) and 84% (noninfected plants) at 50 and 60% fly ash levels, respectively. Fly ash treatments adversely affected root invasion by juveniles, disease intensity and reproduction of the nematodes. A gradual increase in the ash concentration in soil caused a corresponding decrease in the numbers of invading juveniles, galls and egg masses per root system and eggs/egg mass, being lowest at 100% fly ash i.e. 52, 16, 10 and 81 against 289, 137, 131 and 238 (control), respectively. Linear regression suggested 40% fly ash as the most economic level, enhancing yield of infected plants by 96% and suppressing the nematode disease and reproduction by 63 and 76%, respectively.     

南方根结线虫拮抗链霉菌的筛选和鉴定

通过生物测定和温室防病试验 ,对分离自江苏省南京、淮阴和上海三地 3 0份土样的 3 44株放线菌菌株进行筛选 ,得到了 1株对南方根结线虫有较高生物活性的链霉菌菌株LH1 2 1。其培养滤液在离体条件下对南方根结线虫二龄幼虫有较强的触杀作用 ,4h内击倒率达 1 0 0 % ,1 2h内致死率达 1 0 0 % ;番茄植株上的活体防病试验表明 ,2ml/株培养滤液对根结和卵块形成的抑制率分别为52 .8%和 63 .5%。遗传稳定性试验表明LH1 2 1菌株能稳定产生杀线活性物质。LH1 2 1培养滤液主要通过直接触杀二龄幼虫或干扰其侵入植株而达到防治目的。根据形态、培养和生理生化特性的分析 ,将LH1 2 1鉴定为玫瑰暗黄链霉菌     

PGPR对番茄南方根结线虫病的影响

于温室盆栽条件下初步研究了4种根围促生细菌（PGPR）对番茄植株生长和南方根结线虫Meloidogyne incognit病发生情况的影响。结果表明接种多粘芽孢杆菌Bacillus polymyxa或芽孢杆菌菌株B697的番茄植株高度、地上部和地下部干重均显著大于对照,而巨大芽孢杆菌B. megaterium和固氮螺菌Azospirillum sp.菌株A135无促生作用。接种多粘芽孢杆菌＋南方根结线虫和B697菌株＋南方根结线虫两处理的二龄幼虫数、雌虫数、线虫总数、根上卵囊数、卵囊含卵量、发病率和病情指数显著低于只接种南方根结线虫的对照。多粘芽孢杆菌、B697菌株、巨大芽孢杆菌和A135菌株对南方根结线虫的防效分别达65.4%、68.2%、53.8%和53.8%。     

穿刺巴斯德菌活体大量繁殖体系的优化

穿刺巴斯德菌Pasteuria penetrans专性寄生于根结线虫Meloidogyne spp.,具有很好的生防潜力。为了进一步提高其产量,本研究比较了5个对南方根结线虫敏感的番茄栽培品种繁殖穿刺巴斯德菌的效率。结果表明,佳粉18和毛粉802繁殖效率最高,单株番茄上的产孢量分别为1.54×108孢子.株-1和1.47×108孢子.株-1。以佳粉18为根结线虫的寄主植物,研究线虫接种量、接种次数、线虫接入时的苗龄及接种后第1周的培养温度对穿刺巴斯德菌活体繁殖量的影响。研究表明,在番茄9片真叶时,每株番茄接种7500条线虫,分3次接入,根粉的产孢量可提高至1.86×108孢子g-1。在18～31℃温度范围内,温度越高,穿刺巴斯德菌对线虫的侵染率越高;31℃时,其孢子侵染率平均达44.57%。     

Local and Systemic Induced Resistance to the Root-Knot Nematode in Tomato by DL-beta-Amino-n-Butyric Acid

ABSTRACT Chemical inducers of pathogenesis-related proteins and plant resistance were applied to tomato plants, with the aim of inducing resistance to the root-knot nematode Meloidogyne javanica. Relative to control plants, foliar spray and soil-drenching with dl-beta-amino-n-butyric acid (BABA) reduced root-galling 7 days after inoculation, as well as the number of eggs 30 days after inoculation. Other chemicals (alpha- and gamma-amino-n-butyric acid, jasmonic acid, methyl jasmonate, and salicylic acid) were either phytotoxic to tomato plants or did not improve control of root-knot nematodes. Fewer second-stage juveniles invaded BABA-treated tomato roots, and root-galling indices were lower than in control tomato plants. Resistance phenomena in seedlings lasted at least 5 days after spraying with BABA. Nematodes invading the roots of BABA-treated seedlings induced small, vacuolate giant cells. Postinfection treatment of tomato plants with BABA inhibited nematode development. It is speculated that after BABA application tomato roots become less attractive to root-knot nematodes, physically harder to invade, or some substance(s) inhibiting nematode or nematode feeding-site development is produced in roots.     

根结线虫侵染对番茄叶片保护酶活性的影响

通过对番茄感染南方根结线虫后其叶片中几种保护酶活性的变化进行研究。结果表明:接种南方根结线虫后,番茄叶片中的苯丙氨酸解氨酶(PAL)活性即开始低于对照,随后逐渐恢复至对照水平,15 d之后,酶活性又开始显著低于对照水平,一直持续到第30 d,酶活性恢复到对照水平;多酚氧化酶(PPO)活性变化受线虫侵染的影响较小,在试验期间与对照没有显著性差异;超氧化物歧化酶(SOD)的活性在接种后第1 d即显著低于对照水平,至第7 d的时候恢复至对照水平并持续至第30 d。过氧化物酶(POD)活性在1～20 d均显著高于对照水平。因此,POD活性可以作为早期判断番茄是否感染根结线虫的一项指标。     

淡紫拟青霉的培养和对南方根结线虫的防治试验

初步筛选出椰子汁及麦麸为大量繁殖淡紫拟青霉的最适培养基质。室内测定这种真菌对番茄南方根结线虫早期发育阶段卵的寄生率为54％。在室温移栽时,用4克和8克稻谷培养的菌剂处理,50天后线虫卵寄生率分别是35.3％和41.2％。蔬菜塑料大棚小区处理结果表明,移栽前一个月施用这种菌的麦麸加沙培养物,番茄植株平均高度比对照高4～5cm,花果数增加,根结根数和幼虫数量仅为对照的1/3到1/4。     

Relationship between physiological response of French beans of different age to Meloidogyne incognita and subsequent yield loss

A study was made of the effect of a single generation of the root-knot nematode Meloidogyne incognita on the growth of potted French bean plants ( Phaseolus vulgaris ) inoculated at different stages of plant maturity. In separate experiments. 3-, 11- and 13-day-old plants were inoculated before primary leaf expansion (BPLE). at the appearance of trifoliate leaves (TRIF) and at the flower bud (BDS) stages respectively, with 0, 2000, 4000 or 8000 second-stage juvenile nematodes and maintained in a growth chamber under controlled conditions. The photosynthetic rate of the plants inoculated at the TRIF and BDS stages decreased significantly with increasing inoculum level 7 days after inoculation. Although the respiration rate did not significantly change throughout the experimental period, the ratio of photosynthetic to respiration rate decreased significantly with increasing nematode inoculum level and duration of infection. Chlorophyll content, plant dry weight and the numbers of buds, flowers, pods and seeds were significantly lower in infected plants than in the controls; this effect increased with increasing levels of nematode inoculum for all three plant stages. The leaf area was significantly smaller only when nematode infection occurred at the BPLE stage. The plants which were youngest at the time of nematode infection produced the lowest yield; this appeared to result from the effect of nematodes on photosynthesis and related physiological processes.     

温湿度对南方根结线虫卵孵化和二龄幼虫的影响

根结线虫病害是世界上最难防治的蔬菜病害之一,是否可以通过控制生态条件经济有效地防治根结线虫病害值得深入研究。本文利用室内纯培养技术研究了温湿度单因素和双因素对南方根结线虫（Meloidogyne incognita）2龄幼虫(J2)的致死率和卵囊中卵孵化率的影响。试验结果表明,该线虫对温度敏感,40 ℃以上高温和5 ℃以下低温即可抑制J2存活和卵囊中卵孵化,15～30 ℃是线虫的适宜温度;该线虫对湿度不十分敏感,只有干旱（含水量低于1%）和高湿（含水量高于30%）时,对线虫才有一定抑制作用。低温（5 ℃以下低温、含水量5%以上湿度）和高温高湿（35 ℃以上温度和30%含水量）抑制卵囊中卵孵化和J2的存活。本数据的获得将为设施园艺中生态控制南方根结线虫病害提供理论指导和数据支持。     

南方根结线虫Col基因分子克隆及其VIGS效应分析

为了明确线虫角质层胶原蛋白基因(Col)与根结线虫病害的关系,利用从线虫基因组中预测的Col设计特异引物,克隆了南方根结线虫Meloidogyne incognitacol基因MiCol.该基因完整编码区长903bp,编码300个氨基酸.克隆的MiCol与预测的MiCol基因序列一致性高达99.67％,氨基酸序列一致性高达100％.利用病毒介导的基因沉默技术,将其导入番茄植株并接种南方根结线虫,60 d后,沉默载体pTV-MiColi处理番茄植株的根结数比空载体对照及清水对照分别减少49.4％和49.2％,表明MiCol基因沉默显著降低了南方根结线虫的侵染数量.     

Use of crop residues for the control of Meloidogyne incognita under laboratory conditions

This laboratory study evaluates the biofumigant effect of different organic materials with the aim of developing non-chemical alternatives for the management of Meloidogyne incognita (Kofoid & White) Chitwood populations. Sources of organic material from the production system were selected with the aim of reducing agricultural residue accumulation problems as well as decreasing the costs due to the use of chemical fertilizers and pesticides. The selected materials were residues from pepper, strawberry, tomato and cucumber crops, orange juice industry residues, commercial manure and sheep manure, applied at different dosages. Two biofumigation assays were performed under laboratory conditions, using alkaline soils from the Torreblanca area (Murcia, Spain) and acidic soils from the Villa del Prado area (Madrid, Spain). The assays evaluated the effect of the treatments on M. incognita juveniles and other soil organisms, the nematode galling index on tomato roots (susceptible cv. Marmande) grown in the biofumigated soil and soil fertility parameters. The results showed that all biofumigant materials significantly decreased M. incognita populations and galling indices in tomato cv. Marmande. A greater effect was observed on galling indices when applying crop residues together with manure than with the residues alone. Biofumigation had a general beneficial effect on soil fertility, generally increasing nitrogen, organic carbon, pH and potassium levels, and also calcium levels when crop residues of pepper and strawberry were applied. There were no important variations in the number of saprophagous nematodes, dorylaimids and enchytraeids.     

Smoke pollution and the effects of root-knot nematodes on the growth of eggplants

Single and combined effects of smoke pollution and the root-knot nematode, Meloidogyne incognita race 1, were studied on eggplant in 1988 and 1989 at two sites (K1 and K2) 1 and 2 km respectively away from a coal-fired thermal power plant, and a control site at the Department of Botany, Aligarh Muslim University (AMU). Mean concentrations of SO₂, NO₂ and suspended particulate matter (SPM) were respectively 172, 95 and 626 μg/m³ in 1988 and 144, 97 and 556 μg/m³ in 1989 at K1; and 258, 150 and 344 μg/m³ in 1988 and 226, 113 and 293 μg/m³ in 1989 at K2. Peak levels of the gases and SPM were recorded in the early (12.00–15.00) and late (15.00–18.00) afternoon respectively. Concentrations of the pollutants at a control site were very low. Marginal browning appeared on the leaves of eggplants grown at site K2 whether or not they were infected by the nematodes. The galling caused by nematodes was severe at both polluted sites but egg mass production was inhibited at K2. Egg laying was significantly reduced at both polluted sites. M. incognita significantly reduced plant growth, yield and leaf pigment production of eggplant at the control and polluted sites. Most non-infected plants also showed significant reductions in these plant varieties i.e. from the effects of pollution at K2. Foliar concentrations of sulphur were significantly increased at both polluted sites, being greater in infected plants. The interaction of smoke pollution and M. incognita infection led to greater suppressions at the polluted sites, which were mostly significant at K1. At the polluted sites, leaves had more and longer trichomes; also the number and size of stomata were decreased but their openings were wider especially in infected plants at the two polluted sites.     

Protection of olive planting stocks against parasitism of root-knot nematodes by arbuscular mycorrhizal fungi

The effects were investigated, under controlled conditions, of single and joint inoculation of olive planting stocks cvs Arbequina and Picual with the arbuscular mycorrhizal fungi (AMF) Glomus intraradices , Glomus mosseae or Glomus viscosum , and the root-knot nematodes Meloidogyne incognita and Meloidogyne javanica , on plant performance and nematode infection. Establishment of the fungal symbiosis significantly increased growth of olive plants by 88·9% within a range of 11·9–214·0%, irrespective of olive cultivar, plant age and infection by M. incognita or M. javanica . In plants free from AMF, infection by Meloidogyne spp. significantly reduced the plant main stem diameter by 22·8–38·6%, irrespective of cultivar and plant age. Establishment of AMF in olive plants significantly reduced severity of root galling by 6·3–36·8% as well as reproduction of both Meloidogyne spp. by 11·8–35·7%, indicating a protective effect against parasitism by root-knot nematodes. Infection by the nematodes influenced root colonization by AMF, but the net effect depended on the AMF isolate–olive cultivar combination. It is concluded that prior inoculation of olive plants with AMF may contribute to improving the health status and vigour of cvs Arbequina and Picual planting stocks during nursery propagation.     

Pathozone dynamics of Meloidogyne incognita in the rhizosphere of tomato plants in the presence and absence of the nematophagous fungus, Pochonia chlamydosporia

Pathozone dynamics were derived for approx. 50 Meloidogyne incognita juveniles infecting a single root of tomato under controlled conditions from a point source of inoculum and described using simple, non-linear models. The pathozone decayed sigmoidally with distance, but increased over time as progressively more nematodes were able to infect the root. Despite the reported ability of M. incognita juveniles to travel up to 50 cm in some conditions, the maximum width of the pathozone for a single tomato root was estimated as 18·1 mm. It is conjectured that this was because of (i) diffusion from a point source of inoculum, (ii) a small infection court (a single root tip) and (iii) the limited life span of the nematode. A second experiment was used to assess the effect of the nematophagous fungus Pochonia chlamydosporia on the pathozone dynamics of the nematode. This fungus is known to produce nematicidal products in vitro , which may affect invasion of roots by the free-living nematode. To examine the possibility of a change in the position of the site of infection, changes in the probability of gall formation along the root length were also examined. In the absence of P. chlamydosporia , the pathozone dynamics of M. incognita were very similar to those of the first experiment. It was shown that P. chlamydosporia did not significantly affect the pathozone dynamics of M. incognita nor the site of gall formation, which appear to have little importance for the role of the fungus as a biological control agent.     

生物熏蒸结合阳光消毒治理温室根结线虫技术

根结线虫病是危害北方大棚蔬菜的重要病害,严重影响蔬菜的产量。利用“麦秆+石灰氮”及“麦秆+鸡粪+碳酸氢铵”两种方法进行生物熏蒸,结合夏季高温闷棚进行阳光消毒,探讨了对温室蔬菜根结线虫的控制效果。结果表明,生物熏蒸结合阳光消毒措施处理后2个月,根结线虫数量减少40.8%~49.6%、防治效果达60.3%~68.1%;采摘期增产率达18.5%~22.1%,显著高于生防制剂Sr18的控制效果,但低于熏蒸性杀线虫剂棉隆的控制效果。生物熏蒸结合阳光消毒治理蔬菜根结线虫,具有操作简便、防治效果好、投入产出比高等优点,适宜夏季在北方温室蔬菜上使用。     

Cf-2/Rcr3pim番茄品系对南方根结线虫的抗性测定

Cf-2/Rcr3~(pim)基因型番茄不仅能够抵御番茄叶霉菌的侵染,而且对马铃薯金线虫的寄生也有一定的抑制效果。为挖掘根结线虫的新抗性资源,本研究采用室内人工接种法测定了Cf-0/Rcr3~(pim)、Cf-2/Rcr3-3和Cf-2/Rcr3~(pim)基因型番茄品系对南方根结线虫的抗感性。抗性评价结果显示,Cf-0/Rcr3~(pim)品系对南方根结线虫表现高感,Cf-2/Rcr3-3品系为中感,而Cf-2/Rcr3~(pim)品系则为感病。与Cf-0/Rcr3~(pim)和Cf-2/Rcr3-3基因型相比,Cf-2/Rcr3~(pim)基因型番茄品系虽然对南方根结线虫侵染的敏感性略低,但是不能阻止线虫在根系上的大量繁殖,不适于根结线虫的防控应用。