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蔓枯病是危害甜瓜的一种真菌性病害,由于存在生理小种的变异,携带单个抗病基因的甜瓜品种在生产中表现出抗性不足,本研究的目的是评价聚合基因材料的抗性,并分析相关防卫基因的差异表达。利用5份抗源创制8份聚合抗病基因材料,在采用梯度浓度孢子液接种鉴定的基础上,利用RT-PCR技术研究接种后苯丙氨酸解氨酶(PAL)基因、抗坏血酸氧化酶(APX)基因、几丁质酶(CHT)基因等防卫基因在聚合基因材料中的表达情况。结果表明:当接种浓度为5×109mL-1时,单一抗源已开始出现感病现象,而聚合基因材料仍表现高抗或抗,其中,890-398(PI511890×PI482398)和145-471(PI420145×PI140471)抗性最高。890-398中PAL基因表达高峰在1 d,早于PI420145和PI511890的2 d;抗、感及聚合甜瓜材料中的APX基因表达均在接种后3 d达到高峰,890-398中APX基因的表达水平约为对照‘白皮脆’的13.31倍,PI420145和PI511890中APX基因表达水平约为对照的10.00和9.38倍;CHT基因表达最高峰时,890-398中CHT基因的表达量约为对照的28.46倍,PI420145和PI511890约为对照的8.03和20.43倍。结论:甜瓜抗蔓枯病基因的聚合能提高其对蔓枯病的抗性,防卫基因在聚合材料中的高表达或早表达使其表现较高的抗性。本文创制的聚合基因材料可用于甜瓜的抗蔓枯病聚合育种,对甜瓜抗病育种具有重要意义。
As a fungal disease that damages melon, melon blight, due to the presence of physiological races, melon varieties carrying a single disease-resistance gene show insufficient resistance in production. The purpose of this study was to evaluate the resistance of polymeric genetic material , And analyze the differential expression of related defense genes. Based on the identification of the gradient sporophyte inoculum, eight copies of the disease resistance gene were used to establish the gene for the resistance gene, and the effects of PAL gene and ascorbic acid oxidase APX) gene, chitinase (CHT) gene and other defense genes in polymeric gene expression. The results showed that when the inoculation concentration was 5 × 109 mL-1, the single anti-virus began to appear the susceptible phenomenon, but the polymeric gene material still showed high resistance or resistance. Among them, 890-398 (PI511890 × PI482398) and 145-471 PI420145 × PI140471) the highest resistance. The expression of APX gene in resistant, susceptible and polymeric muskmelon reached its peak at 3 days after inoculation, and the expression of APX gene in 890-398 was peaked at 1 day, earlier than PI420145 and PI511890 The level of APX gene expression in PI420145 and PI511890 was about 10.00 and 9.38 times that of the control, respectively. The highest expression of CHT gene was found in CHD gene in 890-398 28.46-fold, PI420145 and PI511890 were about 8.03 and 20.43 times that of the control. CONCLUSION: Polymerization of melon-resistant genes in melon can increase their resistance to villers’ disease. The high or early expression of defense genes in polymeric materials makes them show higher resistance. The polygenetic material prepared in this paper can be used for the anti-griseophytic polymerization breeding of melon, which is of great significance for the disease-resistant breeding of melon.