影响农杆菌介导甜瓜子叶遗传转化的因素

以甜瓜子叶作为根癌农杆菌介导转化的受体,通过GUS基因瞬时表达率的分析,研究此转化体系的最佳实验参数.实验结果表明,预培养时间、预培养后对外植体进行处理、感染时间、共培养时间、农杆菌工程菌的浓度等对转化效率都有一定的影响,但是根癌农杆菌诱导物AS并不能大幅度提高转化效果.对外植体进行重新处理,预培养2 d,用OD560为0.3的农杆菌工程菌感染15～25 min,共培养3～4 d的理想条件下,GUS瞬时表达率可达85%.     

Identification of QTLs related to sugar and organic acid composition in melon using near-isogenic lines

The soluble sugar and organic acid composition of melon (Cucumis melo L.) fruit flesh has become a major focus of plant breeding worldwide in an attempt to improve taste. Thus, sugar and organic acid profiles were characterized using near-isogenic lines (NILs) derived from the Spanish cultivar Piel de Sapo (PS) and the exotic Korean accession Shongwan Charmi (PI 161375). Fruits were cultivated in only one environment. These data were used to map 60 quantitative trait loci (QTLs): 18 for individual sugars, eight for sucrose equivalents, five for the glucose-to-fructose ratio, seven for the total sugar content and 21 for organic acids. Within the QTLs that were associated with the sugar profile, 27 defined the sugar content: eight for fructose, six for glucose, four for sucrose, and nine for sucrose equivalents. Although increased sweetness of selected NILs compared with the parental PS was achieved by an increased glucose or fructose content, only glucose heritability was above 0.5. A total of 21 QTLs (two with positive effects and nineteen with detrimental effects compared with the PS levels) controlled the organic acid profile: l-glutamic, ascorbic and succinic acids (the principal ones) and fumaric, citric, oxalacetic, and isocitric acids. The levels of sugars imparted by the PI 161375 introgression frequently decreased the score grades given to NILs by consumers. Within the 32 QTLs mapped for sensory traits, 27 were associated with lower scores in taste (nine QTLs), sweetness (eight QTLs) or global quality appreciation (nine QTLs); two with increased fruit sourness or sweetness and three with increased fruit bitterness. The QTLs defined herein may assist breeders to understand the overall organoleptic balance (sweetness, sourness, and umami taste) in melon fruit, particularly those located within linkage groups III, V, VI, and VIII to XI.     

Polyamines enhance chilling tolerance of cucumber (Cucumis sativus L.) through modulating antioxidative system

Cucumber chilling-resistant cultivar Changchun mici and -sensitive cultivar Beijing jietou were used in this study to investigate the effects of exogenous PAs on protection against chilling injury as well as on changes of physiological features, and the fluctuation of free PAs content in the leaves under chilling stress. Upon chilling treatment, free spermidine (Spd), spermine (Spm) and putrescine (Put) were remarkably induced in the leaves of cv. Changchun mici 1 day after treatment. The induction of Put declined thereafter, whereas Spd and Spm levels increased steadily. In the leaves of cv. Beijing jietou, Put content was increased only at 1 day after chilling while Spd content decreased significantly upon chilling treatment. Chilling reduced soluble protein content, and decreased the activities of antioxidant enzymes, including superoxidase dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) only in Beijing jietou. However, these changes could be renovated by exogenous application of Put and Spd. It was also found that pretreatment with Put and Spd diminished the increased electrolyte leakage and malondialdehyde (MDA) content caused by chilling in the leaves of both cultivars. Pretreatment of methyglyoxal-bis-(guanylhydrazone) (MGBG), the PAs biosynthetic inhibitor cancelled the effects of PAs in most of the treatments. Moreover, histochemical staining and quantitative measurements showed that exogenous application of Put and Spd eliminated but MGBG exaggerated the hydrogen peroxide (H₂O₂) accumulation caused by chilling stress, especially in leaves of Beijing jietou. Interestingly, Changchun mici was found to contain higher endogenous free PAs contents compared to Beijing jietou. While no significant difference of SOD, POD and CAT activities was found between non-chilling Changchun mici and Beijing jietou seedlings, the former exhibited higher APX activity than the latter. These results suggest that PAs play important roles in the tolerance of cucumber against chilling stress, which is most likely achieved by acting as oxidative machinery against chilling injury.     

甜瓜枯萎病接种方法比较试验初报

分别采用灌根法、米饭粒接种法、蘸根法及胚根接种法对甜瓜枯萎病进行接种试验,拟为甜瓜抗病育种提供依据。结果表明:米饭粒接种法发病率最高,达到99%;胚根接种法最低,为10%;灌根法发病率为66.7%;蘸根法发病率为91%,且操作简单、精确,最适合进行甜瓜枯萎病抗病鉴定研究。     

外源水杨酸诱导的部分甜瓜自毒作用相关基因的表达分析

以甜瓜(Cucumis melo)品种‘新银辉’为材料,采取根灌法,以水杨酸(10μmol/L)处理甜瓜幼苗,通过Real-Time PCR对叶片中部分自毒作用相关基因的表达量进行定量分析,以探究外源水杨酸诱导对甜瓜自毒作用的影响。结果显示,经外源水杨酸诱导后:(1)在催化苯丙烷类代谢途径的3个初始反应的蛋白酶编码基因中,PAL和C4H随处理后时间增加,呈逐渐上调趋势,但PAL比C4H更快响应胁迫,而4CL转录水平没有显著变化;(2)在类黄酮生物合成的3个关键酶的基因中,CHI、F3H均显示出不同程度的下调,CHS于处理后1 d略微上调,然后恢复至处理前水平;(3)两个转录因子基因WD40、R2R3-MYB分别在不同处理时间后显著下调;(4)WD40与CHI的表达相关性较高。据上述结果 ,水杨酸诱导后:(Ⅰ)CHI表达可能与转录因子WD40蛋白相关;(Ⅱ)WD40、R2R3-MYB与PAL、C4H、4CL、CHS间不存在简单、明确的调控关系;(Ⅲ)柚皮苷、表儿茶素、芦丁等甜瓜主要化感物质合成减少。     

不同氮素形态配比对网纹甜瓜干物质分配和氮代谢的影响

该文研究了4种氮素形态配比(NO3--N∶NH4 -N分别为100∶0,75∶25,50∶50和25∶75)对基质栽培网纹甜瓜(品种为“春丽”和“蜜玲珑”)干物质积累和氮代谢的影响。结果表明,不同氮素形态配比影响了植株各器官干重占全株干重的百分比。随氮素形态中氨态氮比例的增加,叶片中硝酸还原酶、硝酸盐含量和可溶性蛋白质含量逐渐降低,而游离氨基酸含量则在NO3--N∶NH4 -N为50∶50的处理中最高。     

AgNO3对甜瓜离体胚囊发育的影响

许多作物的花药、小孢子培养技术被广泛的应用到实际育种工作中。与花药培养相比,离体培养子房或胚珠诱导单倍体植株具有独特的优势,在某些作物上诱导率和绿苗率较高。早在80年代,水稻、大麦、玉米、青稞和马铃薯等都已建立了未受精胚珠单倍体离体培养技术,而对于葫芦科植物至今尚未找到一条高效的单倍体诱导方法。1999年,FICCADENTI进行甜瓜未授粉胚珠离体培养,得到了单倍体植株。2003年,LOTFI采用FICCADENTI的培养方法,对不同甜瓜品种进行诱导,结果只得到了3棵再生植株,在未进行倍性鉴定之前死亡。至今,国内甜瓜离体雌核发育诱导单倍体研究尚未报道。     

直接扩增甜瓜小卫星DNA指纹图谱

以小卫星DNA YNZ22核心序列为引物,甜瓜DNA为模板,在甜瓜上研究了直接扩增小卫星DNA(DAMD)的优化反应体系,结果表明:在20μL的反应体系中,5种主要成分Taq DNA聚合酶、Mg2 、引物、模板DNA和dNTPs的最适浓度分别为1U,2.5 mmol/L,0.5 mmol/L,30 ng,5 mmol/L。在优化的DAMD-PCR反应体系下,利用该引物在28个甜瓜品种上构建了直接扩增小卫星DNA(DAMD)的指纹图谱,分析结果表明,该引物在28个甜瓜品种上共扩增出13位点,其中9位点具有多态性,多态性条带比率为69%,可一次性从28个甜瓜品种中鉴别出其中的21个,鉴别率高达75%。     

甜瓜属新型作物--火参果的研究和利用现状及建议

火参果是我国近年来引进和栽培的新型瓜类水果作物,具有坐果能力强、果实外观漂亮、风味独特、经济效益高和发展前景广阔等优点。就火参果遗传多样性及分类地位、种质资源评价与栽培、抗性及其利用、营养和药用功能成分方面回顾了国内外研究进展,并对火参果栽培技术要点进行总结。此外,由于目前的火参果鲜食品质难以满足大众化的消费需求,市场空间较为有限,制约了火参果产业的进一步发展,提出了解决这个问题相关的建议,旨在为促进我国火参果产业发展提供借鉴。