不同杂草病原真菌的分离及其对野燕麦的致病力

【目的】筛选对野燕麦具有致病作用的生防菌株。【方法】从青海省罹病杂草叶片上分离病原真菌,采用离体生测法和温室盆栽接种法,评价其发酵液对野燕麦的致病性,用盆栽生测法评价其对主栽作物的安全性,并采用形态学结合分子生物学的方法,鉴定其对野燕麦致病力较强的菌株。【结果】分离获得了XN-1、XN-3、XN-4、XN-9和GD-13 5株病原真菌,其中XN-1、XN-3和GD-13的发酵液对野燕麦离体叶片表现出较强的致病力,对种子的萌发抑制作用明显,对野燕麦表现出较强的侵染力,显著抑制植株的生长发育,且对青海省主栽作物中的蚕豆和豌豆相对安全。经鉴定,XN-1、XN-3和GD-13分别为链格孢(Alternariasp.)、黑附球菌(Epicoccum nigium)和短小茎点霉(Phoma exigua)。【结论】XN-1、XN-3和GD-13有望成为防除阔叶田野燕麦的真菌除草剂的候选菌株。     

Control of morningglory species using Fusarium solani and its extracts

Abstract

Natural products of an isolate of Fusarium solani App. & Wr. NRRL 18883 grown on rice medium were discovered to be phytotoxic to morningglory. This isolate was evaluated for biocontrol potential on morningglory species including ivyleaf [Ipomoea hederacea (L.) Jacq.], multicoloured (I. tricolor Cav.), paimleaf (I. wrightii Gray), pitted (I. lacunosa L.), purple moonflower (I. turbinata Lag.), red (I. coccinea L.), sharppod (I. cordatotriloba Dennstedt), smallflower (Jacquemontia tamnifolia (L.) Griseb], and tall [I. purpurea (L.) Roth] morningglory. When sprayed at a concentration of 10 g fungus‐infested rice per 50 ml of water, this isolate caused phytotoxic damage including necrosis, chlorosis, growth inhibition, and mortality. Deleterious effects were recorded in all species, ranging from necrotic spots to death, depending on the species. Mortality ranged from 0% at 3 weeks for purple moonflower to 89% for smallflower morningglory. Soil‐drench application (10–20 ml fungal material per 150 g soil) caused wilting and death. Root lengths of 7‐ and 10‐day‐old plants were reduced between 19 and 89% by soil‐drench application and 17–84% by spray application. Whole plant length reduction of 7‐ and 10‐day‐old plants ranged from 0 to 96%, and 39 to 96% for soil‐drench and spray applications, respectively. Testing of F. solani on weed and crop species showed that most broadleaved species were sensitive but monocotyledons were immune. Tests of the phytotoxic extract for known phytotoxins such as common trichothecene (deoxynivalenol) and non‐trichothecene compounds (fusaric acid, moniliformin, fumonisins) were negative. This is the first report of the use of F. solani against morningglory.     

Infection of Nicotiana species by the anthracnose fungus, Colletotrichum orbiculare

Colletotrichum gloeosporioides f. sp. malvae, isolate Biomal®, ATCC 20767, was originally isolated from round-leaved mallow (Malva pusilla) and developed as a weed biocontrol agent. Ribosomal DNA sequence analysis was recently used to re-classify this fungus as C. orbiculare, which is an aggregate species with a number of formae speciales. Several morphological features of ATCC 20767 were examined that were consistent with those described for C. orbiculare, and inoculation of a number of Nicotiana species and several cultivars of N. tabacum showed that this fungus was pathogenic to many of these previously undescribed hosts. Spore germination and appressorium formation were higher on tobacco than previously observed on round-leaved mallow. The pathogen produced melanized appressoria on N. tabacum leaves that formed preferentially at the anticlinal epidermal cell wall. A symptomless phase of infection persisted for 72–96 h postinoculation, during which time the fungus first produced a spherical infection vesicle from an infection peg, and then large primary hyphae which grew through the epidermal cells. The large primary hyphae were highly constricted at the points of penetration of the host cell walls. Thin secondary hyphae appeared at 96–120 h postinoculation coinciding with the appearance of light green, water-soaked spots and the formation of acervuli. The infection of tobacco by C. orbiculare ATCC 20767 is not a non-specific interaction but appears to follow an intracellular hemibiotrophic infection process that is very similar to that established for the C. orbiculare infection of round-leaved mallow, cucurbits and beans.     

Relationship between aggressiveness of <Emphasis Type="Italic">Stagonospora </Emphasis>sp. isolates on field and hedge bindweeds,and <Emphasis Type="Italic">in vitro</Emphasis> production of fungal metabolites cercosporin,elsinochrome A and leptosphaerodione

Stagonospora convolvuli LA39, an effective biocontrol agent of Convolvulus arvensis (field bindweed) and Calystegia sepium (hedge bindweed) produces phytotoxic metabolites leptosphaerodione and elsinochrome A. Stagonospora isolate 214Caa produces the toxin cercosporin. If toxic metabolite production is not linked to the pathogenic ability of the fungus on bindweeds, selection of aggressive strains with limited or no production of the metabolites would reduce any perceived risk of using strains of the fungus as a mycoherbicide. Therefore, 30 isolates of Stagonospora sp. including LA39 and 214Caa were characterised for aggressiveness on both bindweeds, and production of the three metabolites. Nine isolates were more aggressive than LA39 on both bindweeds. Classification of isolates based on metabolite type agreed largely with previous similar characterisation based on polymerase chain reaction-restriction fragment length polymorphism of internal transcribed spacer of ribosomal DNA. Cercosporin producers produced neither leptosphaerodione nor elsinochrome A and together with isolates that produce none of the three metabolites, were less pathogenic on bindweeds. Conversely, there was a positive correlation between elsinochrome A and leptosphaerodione production, and each was positively correlated with aggressiveness of isolates on both bindweeds. Generally, any isolate where elsinochrome A was not detected was not aggressive on any of the two bindweeds. This probably implies that selecting elsinochrome A-negative, but aggressive Stagonospora strain(s) may be difficult. However, aggressive isolates may not produce elsinochrome A in planta at levels that could constitute any risk in the environment. In a preliminary attempt to determine the levels of elsinochrome A and leptosphaerodione produced in diseased bindweeds, none of the toxins was detected in Stagonospora infected bindweed leaves. Detailed investigation focusing on the detection and quantification of in planta production of elsinochrome A by Stagonospora isolates, and determination of the fate of elsinochrome A in the environment, and its relationship with leptosphaerodione may be essential. Similarly, development of molecular tools to monitor the mycoherbicide following field application is vital.