Isolation of Pathogenicity Mutants of Fusarium oxysporum f. sp. melonis by Insertional Mutagenesis

Restriction enzyme-mediated integration (REMI) mutagenesis was used to isolate mutants of Fusarium oxysporum f. sp. melonis impaired in pathogenicity. The race 2 strain Mel02010 was transformed with linearized pSH75, conferring resistance to hygromycin B, with or without the enzyme used to linearize the plasmid. Addition of restriction enzymes did not affect the transformation frequency. A total of 2929 REMI transformants were tested for pathogenicity to three melon cultivars, Amus, Ogon 9 and Ohi. The race 2 strains are pathogenic to Amus and Ogon 9, but not to Ohi. Of 43 transformants with reduced pathogenicity on susceptible melon cultivars, 12 mutants were examined in detail for pathogenicity, vegetative growth and integrative mode of pSH75. The levels of pathogenicity varied among these mutants. Two mutants (B48 and B137) almost completely lost pathogenicity to both susceptible cultivars, and the others had reduced pathogenicity. Mutants B48, B241, B886 and X36 were also impaired in vegetative growth. Mutant B809 was a biotin auxotroph. By DNA gel blot analysis, nine mutants were found to contain a single copy of the transformation vector. These mutants may thus be useful in isolating genes involved in pathogenicity. Received 22 December 2000/ Accepted in revised form 16 April 2001     

几种杀菌剂对甜瓜枯萎病菌的室内毒力测定

采用生长速率法和抑菌圈法测定了9种杀菌剂对甜瓜枯萎病菌(Fusarium oxysporum f.sp.melonis)的毒力。抑菌圈法测定结果表明多菌灵和咯菌腈对甜瓜枯萎病菌的毒力最高,其3d的EC50分别为783.7905mg·L-1和975.2145mg·L-1;生长速率法测定结果表明:多菌灵和多菌灵-五氯硝基苯对病原菌的抑制率最高,7d的EC50分别为2.4757mg·L-1和3.3570mg·L-1。     

快速鉴定瓜类疫病抗性方法的建立及黄瓜种质资源鉴定

【目的】为方便研究瓜类疫病,快速筛选抗疫病的瓜类材料,建立快速安全有效的瓜类疫病抗性鉴定体系。【方法】通过灌根、喷雾、浸根、菌块离体接种等4种接种方法对黄瓜抗病材料和感病材料幼苗进行接种,通过比较发病率、发病时间及操作程序等确定最简便有效的鉴定方法。【结果】4种接种方法均能鉴定黄瓜疫病抗感情况,但是鉴定效果差异显著。浸根法、喷雾法、灌根法接种合适浓度均为10~2个孢子/mL,操作复杂,接种效率低。菌块离体接种方法利用2 mm直径菌块直接接种于离体子叶背面,放置于有润湿滤纸的培养皿中,保湿24 h即可观察表型,通过计算病斑大小快速鉴定瓜类材料的抗性;利用该方法对40份不同抗性的黄瓜材料进行鉴定,发现栽培黄瓜抗疫病材料极少,只有PE40、PE119、PE120、PE185等4份材料具有抗性,90%的材料均为感病或高感。【结论】菌块子叶离体接种法是瓜类疫病鉴定的快捷有效方法,该方法无需诱孢,能够一次性鉴定数百份材料,且能够保存感病材料或中间材料,大大提高鉴定效率,为瓜类疫病抗性育种奠定基础。     

几种新型杀菌剂对瓜类蔓枯病菌的毒力测定

用生长速率法测定8种药剂对瓜类蔓枯病的抑制作用表明：30％苗菌敌抑制作用最强,40mg／kg抑制率为100％,其次为50％扑海因和5％菌毒清,600mg／kg抑制率分别为914％和837％;用最小二乘法计算的3种药剂的毒力回归方程依次为：Y＝4．2187＋2．1317x、Y＝0．8644＋2．2214x、Y＝3．6955＋0．9434x;EC50为2．3255mg／kg、73．378mg／kg、24．1474mg／kg;EC95为14．342mg／kg．410．964mg／kg。1338．mg／kg为田间进一步试验提供了理论依据。     

Occurrence of Fusarium oxysporum f. sp. melonis Race 1 in Japan

In 1994, Fusarium wilt of melon cultivars which are resistant to races 0 and 2 of Fusarium oxysporum f. sp. melonis was observed in southern area of the Lake Biwa region, Shiga prefecture. In commercial fields, mature plants of cv. Amus which were grafted onto cv. Enken Daigi 2, and of cv. FR Amus showed yellowing, wilting and finally death before harvesting of fruits. Diseased plants had vascular and root discolorations, and their stem sections yielded typical colonies of F. oxysporum. When the Shiga strains were tested for their pathogenicity to 12 species of cucurbits, they caused wilts only on melon. Using race differential cultivars of melon, the Shiga strains were classified as race 1 of F. oxysporum f. sp. melonis, which has not been reported in Japan. To further characterize their pathogenicity, the strains were used to inoculate 46 additional cultivars of melon, oriental melon and oriental pickling melon. All the race 1 strains were pathogenic to the cultivars tested, and their host range was apparently different from those of strains belonging to other races (races 0, 2 and 1,2y). DNA fingerprinting with a repetitive DNA sequence, FOLR3, differentiated race 1 strains from strains of races 0 and 2, but not from race 1,2y strains. Received 2 July 1999/ Accepted in revised form 30 September 1999     

利用诱导因子抗新疆甜瓜疫霉病的田间效果

用化学诱导因子和灭活的生物诱导因子在田间自然条件下诱导了新疆甜瓜产生对疫霉病的整体抗性。1996年的试验结果表明，用化学诱导因子诱导处理，最高诱抗效果可达79．40％，平均诱抗效果在40％以上；生物诱导因子诱导处理，最高诱抗效果为68．2％，平均诱抗效果为30％以上。1997年的试验基本上验证了1996年的结果，化学诱导因子最高诱抗效果可达60％，平均诱抗效果为35．86％，经t测验分析，诱抗效果显著。     

Characterization of Fusarium oxysporum f. sp. radicis-cucumerinum attacking melon under natural conditions in Greece

A severe root and stem rot disease of melon was observed during the 2001 growing season on four glasshouse crops in Heraklio, Greece. A total of 43 isolates of F. oxysporum , obtained in Crete from glasshouse-grown melon and showing fusarium wilt or root and stem rot symptoms, were characterized by pathogenicity and vegetative compatibility. The majority of these isolates was also fingerprinted via amplified fragment length polymorphic (AFLP) analysis. Of the total number of isolates, 22 were identified by pathogenicity tests as F. oxysporum f. sp. melonis , 20 as F. oxysporum f. sp. radicis-cucumerinum , while one isolate was nonpathogenic on cucumber, melon, sponge gourd and pumpkin. All 22 isolates of F. oxysporum f. sp. melonis were assigned to vegetative compatibility group (VCG) 0134, and all 20 isolates of F. oxysporum f. sp. radicis-cucumerinum to VCG 0260. Isolates of F. oxysporum f. sp. radicis-cucumerinum were incompatible with isolates of F. oxysporum f. sp. melonis. AFLP fingerprinting allowed for the clustering of the isolates of the two formae speciales of F. oxysporum along two separate phenetic groups: f. sp. melonis to AFLP major haplotype I, and f. sp. radicis-cucumerinum to AFLP major haplotype II. Overall, pathogenicity, vegetative compatibility grouping and AFLP analysis were correlated and effectively distinguished isolates of F. oxysporum from melon. This appears to be the first report of natural infection of melon by F. oxysporum f. sp. radicis-cucumerinum worldwide.     

防治甜瓜枯萎病药剂配方筛选及盆栽防效测定

以甜瓜枯萎病菌为试材,采用菌丝生长速率法和孢子萌发法从8种待选药剂中选出4种较优试剂进行复配试验,采用Wadley方法分别计算菌丝生长和孢子萌发的增效系数,确定最优配比并进行盆栽试验。结果表明:8种待选药剂中,25%腈菌唑乳油和45%咪鲜胺微乳剂对菌丝生长抑制效果较好,EC50值分别为0.001 2、0.004 6 mg·L^-1;250 g·L^-1嘧菌酯悬浮剂和25%啶菌恶唑乳油对孢子萌发抑制效果较好,EC50分别为0.092 2、3.263 2 mg·L^-1。上述4种药剂按比例法两两混配,30种组合中有2种配比对病菌菌丝生长及孢子萌发均为增效和加和作用,即咪鲜胺∶嘧菌酯=4∶1,嘧菌酯∶啶菌恶唑=2∶3。盆栽接种防效测定结果表明,上述2个组合每株浇灌10 mL浓度为1.0 g·L^-1药液对甜瓜枯萎病防治效果分别为80.42%和76.57%,均显著高于单剂防效。为甜瓜枯萎病的防治药剂使用提供参考依据。     

内吸性杀菌剂对甜瓜枯萎病菌的室内毒力测定

采用生长速率法测定了几种内吸性杀菌剂对甜瓜枯萎病菌的毒力.试验结果表明,多菌灵和氟硅唑的毒力最高,其EC50分别为2.267 1 mg/L和13.341 8 mg/L;而且枯萎病菌对多菌灵和氟硅唑的敏感性也较高;多菌灵和氟硅唑时枯萎病菌的联合毒力测定结果表明,两种药以6:1和1:6混配,具有增效作用.     

黄瓜蔓枯病菌生物学特性及室内药剂筛选

研究了黄瓜蔓枯病菌(Mycosphaerellamelonis)生物学特性,并对药剂进行了室内初步筛选。结果表明:该菌菌丝生长的适宜温度范围为20～30℃,最适温度为25℃。分生孢子萌发的最适温度为30℃,最适pH值为6,致死温度为45.5℃10min。全黑暗和全光照处理下的菌落直径与12h光暗交替在P<0.01时差异极显著。病菌对C源营养的利用率高于N源,其中对乳糖的利用率最高。供试的6种药剂中以15%三唑酮可湿性粉剂对病菌的抑制作用最强。