黄瓜嫁接砧木的筛选及亲和性机理研究
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摘要
黄瓜(Cucumis sativus L.)是一种重要的瓜类蔬菜,其嫁接育苗技术已广泛地应用于实际生产。黄瓜砧木品种不是很丰富,而嫁接亲和性机理的研究大多停留在生理生化方面的研究,在分子生物学方面的研究较少。本研究以津春四号黄瓜为接穗,南瓜、瓠子为砧木,以津春四号/黑籽南瓜嫁接苗,以及津春四号自根苗为对照,来筛选砧木品种;利用中国南瓜57自嫁苗、津春四号黄瓜自嫁苗和津春四号/中国南瓜嫁接苗的植株性状和组织切片,从个体水平和细胞水平来研究嫁接苗嫁接亲和性机理;以中国南瓜57自嫁苗、津春四号黄瓜自嫁苗以及津春四号/中国南瓜57嫁接苗,嫁接后不同时期愈合部位显微结构观察以及mRNA差异表达分析,从细胞水平和分子水平来研究嫁接苗共生亲和性机理。实验主要结果如下:
1.砧木筛选,接穗为津春四号黄瓜,砧木为实验室自选的瓠子华瓠杂3号、中国南瓜品系45、57、78、80,对照为黑籽南瓜做砧木的嫁接苗,嫁接后统计其成活率,调查从定植后第五天开始,每隔五天记载嫁接苗植株性状,及相关农艺性状,同时测量品质,对照为黑籽南瓜做砧木的嫁接苗以及津春四号自根黄瓜。结果表明:试验所用的砧木嫁接后成活率均在90%以上,华瓠杂3号嫁接苗的产量极显著的高于黑籽南瓜、中国南瓜品系做砧木的嫁接苗以及黄瓜自根苗。定植后25天,黑籽南瓜和华瓠杂3号嫁接苗以及黄瓜自根苗在接穗茎粗、砧木茎粗、主蔓长、叶片数、叶片长和叶片宽等性状与中国南瓜品系间存在显著差异。华瓠杂3号的第一雌花节位较低,与黑籽南瓜、中国南瓜品系间存在极显著差异,但与自根黄瓜间无显著差异,华瓠杂3号黄瓜嫁接苗的根瓜节位极显著低于黄瓜自根苗和其它砧木黄瓜嫁接苗。农艺性状的聚类分析结果显示,华瓠杂3号、黑籽南瓜、黄瓜自根苗聚为一类。不同砧木嫁接黄瓜品质无显著差异。说明华瓠杂3号做砧木并没有影响黄瓜的生长,但是显著提高了黄瓜的产量,是黄瓜丰产栽培的好的砧木品种。
2.嫁接亲和性机理的研究主要从以下2个方面进行:嫁接亲和力分析,通过中国南瓜57自嫁苗、津春四号黄瓜自嫁苗以及津春四号/中国南瓜57嫁接苗嫁接后不同时期愈合部位组织结构观察以及基因差异表达分析,结果表明:津春四号黄瓜自嫁苗的愈合速度快于中国南瓜57自嫁苗以及津春四号/中国南瓜57嫁接苗。不同时期南瓜自嫁苗愈合部位的cDNA-AFLP结果分析表明,特异性带均出现在嫁接后第6天取样的材料中。共生亲和力分析,以华瓠杂3号、黑籽南瓜和中国南瓜57做砧木,接穗为津春四号黄瓜。定植后调查不同时期植株性状、研究嫁接苗伤流液对黄瓜生长的影响以及嫁接苗愈合部位的组织形态观察;结果表明:不同砧木嫁接接穗植株生长情况有差异,趋势表现为黑籽南瓜和华瓠杂3号的长势要较中国南瓜57强;华瓠杂3号做砧木的嫁接苗伤流液对黄瓜苗的生长有明显的促进作用;嫁接苗愈合部位的组织结构观察显示,华瓠杂3号做砧木的嫁接苗维管束比黑籽南瓜和中国南瓜57的维管束发达。说明嫁接亲和力强的嫁接组合植株长势较旺盛,维管束发达,且产生促进接穗生长的激素。
本试验砧木筛选筛选出的黄瓜嫁接砧木华瓠杂3号可以广泛的应用于黄瓜嫁接上,嫁接亲和机理的研究为以后的嫁接亲和性的研究提供一定的理论依据。
Cucumber (Cucumis sativus L. ) is a important gourd vegetable, the technology of graft breeding of cucumber is widely used to practical production. The rootstocks of cucumber is not enough, study of the mechanisms of affinity on cucumber almost rest on the aspects of physiology and biochemistry. This experiment used JinChun-4 as scion and pumpkin, bottle gourd as rootstock, the grafted seedlings of JinChun-4/fig-leaf-gourd and self-rooted cucumber seedlings as CK, selected rootstock with good affinity. Microstructure measurement and mRNA differential expression of healing parts at different stages of the grafted seedlings of self-grafteded Chinese pumpkin 57, self-grafted JinChun-4 and JinChun-4/fig-leaf-gourd were used to study the mechanism of graft affinity from the levels of cells and molecular. This experiment used plant characters and tissue sections of JinChun-4/Chinese pumpkin 57, JinChun-4/bottle gourd Hua No.3 and JinChun-4/fig-leaf-gourd, studied the mechanisms of symbiotic affinity from individual and cell levels. The main results were as follows:
1. Rootstock screening, the survival rate of grafting was statisticsed with JinChun-4 as scion and pumpkin(Chinese pumpkin strains 45、57、78 and 80), bottle gourd (bottle gourd Hua No.3) as rootstock, the grafted Seedlings of JinChun-4/fig-leaf-gourd and self-rooted cucumber seedlings as CK, the fifth day after planting characters of grafted cucumber seedlings were surveyed at the first time, then every five days surveyed once, yield were investigated and agronomic characters were recorded, quality of merchantable cucumber fruit were measured. The grafted seedlings of JinChun-4/fig-leaf-gourd and self-rooted cucumber seedlings were control. The results showed that, surviving rate of different grafted plants is above 90%, yield of bottle gourd Hua No.3 was significant higher fig-leaf-gourd, Chinese pumpkin strains and self-rooted cucumber. The twenty-fifth day after planting, fig-leaf-gourd, bottle gourd Hua No.3 and self-rooted cucumber had no obvious differences in scion widen, stem widen, main vine height, leaf number, leaf length and wide leaf blade, but had significant differences with Chinese pumpkin strains. The node of 1st female flower of bottle gourd Hua No.3 was significant lower than fig-leaf-gourd and Chinese pumpkin strains, but was no obvious difference with self-rooted cucumber. Bottle gourd Hua No.3 has the lowest node of root-frut among stocks. Grafting had no obvious effects on fruit quality. Fig-leaf-gourd, Bottle gourd Hua No.3 and self-rooted cucumber can be classified into one group by cluster analysis of characters. These results indicated that bottle gourd Hua No.3 had no obvious effects on the growth of cucumber, but significant improved the yield of cucumber, bottle gourd Hua No.3 were good stocks of cucumber.
2. The study of understanding the mechanism of grafting compatibility was in this way: Microstructure measurement and mRNA differential expression of healing parts at different stages of the grafted seedlings of self-grafted Chinese pumpkin 57, self-grafted JinChun-4 and JinChun-4/fig-leaf-gourd were used to study the graft affinity. The results showed that healing speed of the grafted Seedlings of Self-grafted JinChun-4 was faster than others. The result of cDNA-AFLP of pumpkin grafting seedling which rootstock was itself at different phase showed the specific bands would appear in samples after grafting 6 days. Chinese pumpkin 57, bottle gourd Hua No.3 and fig-leaf-gourd as rootstock, JinChun-4 as scion were used to research symbiotic affinity. After planting, characters of grafting seeding were investigated at different phases, the effects of xylem sap about cucumber growth was studied, and the healing parts of grafting seeding were observed in anatomical morphology. The results showed that the effects of different rootstocks were different. The trend of scion growth was fig-leaf-gourd, bottle gourd Hua No.3 as rootstock was better than Chinese pumpkin 57. The xylem sap of bottle gourd No.3 as rootstock was the best one to promote grafting seedling of cucumber to grow significantly. The results of anatomical morphology of the healed sections of grafting seeding showed vascular bundle of grafting seeding which bottle gourd Hua No.3 was as rootstock was the best developed than others. These results indicated that grafted combination with good affinity has developed vascular and produces hormones which promoted the growth of plants.
This research screened a good rootstock of cucumber named bottle gourd Hua No.3 which can be used to graft of cucumber, studied the mechanism of affinity on graft which can provide theoretical basis for the study of the mechanism of affinity.
1.砧木筛选,接穗为津春四号黄瓜,砧木为实验室自选的瓠子华瓠杂3号、中国南瓜品系45、57、78、80,对照为黑籽南瓜做砧木的嫁接苗,嫁接后统计其成活率,调查从定植后第五天开始,每隔五天记载嫁接苗植株性状,及相关农艺性状,同时测量品质,对照为黑籽南瓜做砧木的嫁接苗以及津春四号自根黄瓜。结果表明:试验所用的砧木嫁接后成活率均在90%以上,华瓠杂3号嫁接苗的产量极显著的高于黑籽南瓜、中国南瓜品系做砧木的嫁接苗以及黄瓜自根苗。定植后25天,黑籽南瓜和华瓠杂3号嫁接苗以及黄瓜自根苗在接穗茎粗、砧木茎粗、主蔓长、叶片数、叶片长和叶片宽等性状与中国南瓜品系间存在显著差异。华瓠杂3号的第一雌花节位较低,与黑籽南瓜、中国南瓜品系间存在极显著差异,但与自根黄瓜间无显著差异,华瓠杂3号黄瓜嫁接苗的根瓜节位极显著低于黄瓜自根苗和其它砧木黄瓜嫁接苗。农艺性状的聚类分析结果显示,华瓠杂3号、黑籽南瓜、黄瓜自根苗聚为一类。不同砧木嫁接黄瓜品质无显著差异。说明华瓠杂3号做砧木并没有影响黄瓜的生长,但是显著提高了黄瓜的产量,是黄瓜丰产栽培的好的砧木品种。
2.嫁接亲和性机理的研究主要从以下2个方面进行:嫁接亲和力分析,通过中国南瓜57自嫁苗、津春四号黄瓜自嫁苗以及津春四号/中国南瓜57嫁接苗嫁接后不同时期愈合部位组织结构观察以及基因差异表达分析,结果表明:津春四号黄瓜自嫁苗的愈合速度快于中国南瓜57自嫁苗以及津春四号/中国南瓜57嫁接苗。不同时期南瓜自嫁苗愈合部位的cDNA-AFLP结果分析表明,特异性带均出现在嫁接后第6天取样的材料中。共生亲和力分析,以华瓠杂3号、黑籽南瓜和中国南瓜57做砧木,接穗为津春四号黄瓜。定植后调查不同时期植株性状、研究嫁接苗伤流液对黄瓜生长的影响以及嫁接苗愈合部位的组织形态观察;结果表明:不同砧木嫁接接穗植株生长情况有差异,趋势表现为黑籽南瓜和华瓠杂3号的长势要较中国南瓜57强;华瓠杂3号做砧木的嫁接苗伤流液对黄瓜苗的生长有明显的促进作用;嫁接苗愈合部位的组织结构观察显示,华瓠杂3号做砧木的嫁接苗维管束比黑籽南瓜和中国南瓜57的维管束发达。说明嫁接亲和力强的嫁接组合植株长势较旺盛,维管束发达,且产生促进接穗生长的激素。
本试验砧木筛选筛选出的黄瓜嫁接砧木华瓠杂3号可以广泛的应用于黄瓜嫁接上,嫁接亲和机理的研究为以后的嫁接亲和性的研究提供一定的理论依据。
Cucumber (Cucumis sativus L. ) is a important gourd vegetable, the technology of graft breeding of cucumber is widely used to practical production. The rootstocks of cucumber is not enough, study of the mechanisms of affinity on cucumber almost rest on the aspects of physiology and biochemistry. This experiment used JinChun-4 as scion and pumpkin, bottle gourd as rootstock, the grafted seedlings of JinChun-4/fig-leaf-gourd and self-rooted cucumber seedlings as CK, selected rootstock with good affinity. Microstructure measurement and mRNA differential expression of healing parts at different stages of the grafted seedlings of self-grafteded Chinese pumpkin 57, self-grafted JinChun-4 and JinChun-4/fig-leaf-gourd were used to study the mechanism of graft affinity from the levels of cells and molecular. This experiment used plant characters and tissue sections of JinChun-4/Chinese pumpkin 57, JinChun-4/bottle gourd Hua No.3 and JinChun-4/fig-leaf-gourd, studied the mechanisms of symbiotic affinity from individual and cell levels. The main results were as follows:
1. Rootstock screening, the survival rate of grafting was statisticsed with JinChun-4 as scion and pumpkin(Chinese pumpkin strains 45、57、78 and 80), bottle gourd (bottle gourd Hua No.3) as rootstock, the grafted Seedlings of JinChun-4/fig-leaf-gourd and self-rooted cucumber seedlings as CK, the fifth day after planting characters of grafted cucumber seedlings were surveyed at the first time, then every five days surveyed once, yield were investigated and agronomic characters were recorded, quality of merchantable cucumber fruit were measured. The grafted seedlings of JinChun-4/fig-leaf-gourd and self-rooted cucumber seedlings were control. The results showed that, surviving rate of different grafted plants is above 90%, yield of bottle gourd Hua No.3 was significant higher fig-leaf-gourd, Chinese pumpkin strains and self-rooted cucumber. The twenty-fifth day after planting, fig-leaf-gourd, bottle gourd Hua No.3 and self-rooted cucumber had no obvious differences in scion widen, stem widen, main vine height, leaf number, leaf length and wide leaf blade, but had significant differences with Chinese pumpkin strains. The node of 1st female flower of bottle gourd Hua No.3 was significant lower than fig-leaf-gourd and Chinese pumpkin strains, but was no obvious difference with self-rooted cucumber. Bottle gourd Hua No.3 has the lowest node of root-frut among stocks. Grafting had no obvious effects on fruit quality. Fig-leaf-gourd, Bottle gourd Hua No.3 and self-rooted cucumber can be classified into one group by cluster analysis of characters. These results indicated that bottle gourd Hua No.3 had no obvious effects on the growth of cucumber, but significant improved the yield of cucumber, bottle gourd Hua No.3 were good stocks of cucumber.
2. The study of understanding the mechanism of grafting compatibility was in this way: Microstructure measurement and mRNA differential expression of healing parts at different stages of the grafted seedlings of self-grafted Chinese pumpkin 57, self-grafted JinChun-4 and JinChun-4/fig-leaf-gourd were used to study the graft affinity. The results showed that healing speed of the grafted Seedlings of Self-grafted JinChun-4 was faster than others. The result of cDNA-AFLP of pumpkin grafting seedling which rootstock was itself at different phase showed the specific bands would appear in samples after grafting 6 days. Chinese pumpkin 57, bottle gourd Hua No.3 and fig-leaf-gourd as rootstock, JinChun-4 as scion were used to research symbiotic affinity. After planting, characters of grafting seeding were investigated at different phases, the effects of xylem sap about cucumber growth was studied, and the healing parts of grafting seeding were observed in anatomical morphology. The results showed that the effects of different rootstocks were different. The trend of scion growth was fig-leaf-gourd, bottle gourd Hua No.3 as rootstock was better than Chinese pumpkin 57. The xylem sap of bottle gourd No.3 as rootstock was the best one to promote grafting seedling of cucumber to grow significantly. The results of anatomical morphology of the healed sections of grafting seeding showed vascular bundle of grafting seeding which bottle gourd Hua No.3 was as rootstock was the best developed than others. These results indicated that grafted combination with good affinity has developed vascular and produces hormones which promoted the growth of plants.
This research screened a good rootstock of cucumber named bottle gourd Hua No.3 which can be used to graft of cucumber, studied the mechanism of affinity on graft which can provide theoretical basis for the study of the mechanism of affinity.
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46.于贤昌,刑禹贤.黄瓜嫁接苗抗冷特性研究.园艺学报,1997,24(4):348-352.
47.于贤昌等.蔬菜嫁接的研究与应用.山东农业大学学报,1998,29(2):249-256.
48.于贤昌,邢禹贤.不同砧木与接穗对黄瓜嫁接苗抗冷性的影响.中国农业科学,1998,31(2):41-47.
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50.喻景权,杜尧舜.蔬菜设施栽培可持续发展中的连作障碍问题.沈阳农业大学学报,2000,31(1):124-126.
51.曾义安,朱月林,黄保健,等.黑籽南瓜砧木对黄瓜生长结实、抗病性及营养元素含量的影响.植物资源与环境学报,2004,13(4):15-19.
52.曾义安,朱月林,黄保健,等.嫁接黄瓜光和特性及叶片激素含量和可溶性蛋白研究.南京农业大学学报,2005,28(1):16-19.
53.张红梅,金海军,余纪柱,等.不同南瓜砧木对嫁接黄瓜生长和果实品质的影响.内蒙古农业大学学报,2007,28(3):177-181.
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57.赵青青.嫁接对茄子生长发育和黄萎病抗性的影响.中国蔬菜,1997,6:7.
58.周宝利,高艳新.嫁接茄子抗病性和电导率、脯氨酸含量及苯丙氨酸酶活性的关系.园艺学报.1998a,25(3):300-302
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60.周宝利,王月英.以“托鲁巴姆”为砧木的茄子嫁接苗耐低温特性研究.辽宁农业科学,1999(1):5-8.
61.周宝利,林桂荣,高新艳等.嫁接茄子黄姜病抗性与苯丙烷类代谢的关系.沈阳农业大学学报,2000,31(1):57-60.
62.周长勇,张秀清.番茄嫁接苗与自根苗的对比试验.中国蔬菜,2001,(4):32-33.
63.祖容,于泽源.三种葡萄砧木解剖构造与嫁接植株新梢生长关系.北方园艺,1998(10):12-14
64.Arian Dijkhuizen;Jack E.Staub.QTL conditioning yield and fruit quality traits in cucumber:effects of environment and genetic background.Journal of new seeds,2002,14(4):1-3.
65.Breyne P,Zabeau M.Genome-wide expression analysis of plant cell cycle modulated genes.Current Opinion in Plant Biology,2001,4:136-142.
66.Ditt RF,Nester EW,Comai L.Plant gene espression response to Agrobacterrium tumefaciens.Proceedings of the National Academy of Sciences.USA,2001,4:136-142.
67.Heo Y C.Effects of rootstocks on exudation and mineral elements contents indifferentparts of oriental melon ancucumber(in Korean with English summary ).MS thesis,Kyung Hee University,Seoul,Korea,1991,53.
68.Gebologlu N;Saglam The effect of different plant spacing and mulching materials on the yield and fruit quality of pickling cucumber.Acta Horticulturae,2002,57(9): 603-607.
69. GorrisL GM, Smid EJ. Control of fongal storage diseases of potato by use of plant-essential oil components. Brighton Crop Proteetion Conference Pest and Diseases, 1994, 1: 307-312.
70. Jeffree C. E. Development of intercellular connections between opposing cells in graft union. The New Physiologist, 1983, 93: 491.
71. Hong Chen, Yong-Qing Wang. Genetic variation in the grafting union of tomato and eggplant. American-Eurasian Journal of Agricultural and Environmental Sci, 2006,1(1): 37-41.
72. Hossain MS, Ferdous AJ, Hasan CM. Invitro antimicrobial activities of alkaloids from the stem Bark of Desmos longiflorus Roxb. Bangladesh Journal of Botany,1993, 22(1): 34-40.
73. Kato T, Lou H. Effect of rootstocks on the yield, mineral nutrition and hormone level in xylem sap in eggplant. Journal of Japan Society Sc, 1989, 58(2): 345-352.
74. Vos Pieter. Rene Houppe AFLP; A new technique for DNA fingerprinting. Nucleic Acids Research, 1995, 23(21): 4407-4414.
75. Singh uP, Paithiviraj B, Wagner KG, et al. Effeet of ajoene, a consrituent of garlic(Alliumsativum)on powdery mildew of pea(Pisumsativum). Journal of Plant Diseases and Protection, 1995, 102: 399-406.
76. Rosa M R, Juan M R, Luis R. Role of grafting in horticulture plants under stress conditions. Food, Agriculture Environment, 2003, 1(1): 70-74.
77. Stuart J, Harrison, Kenneth C, et al. An antimierobial peptide from the australia native hardenbergia violacea provides the first funetionally characterized membe of asub family of plant defensins. Plant Physiology, 1997, 24: 571-578.
78. Ross SA, Megalla SE, Bishay DW, et al. Studies for determining antibiotie substances in some egy ptian plants partll. Antimierobi alalkaloids from the seeds of peganum harmala FitoteraPia, 1990, 15(6): 309-312.
79. Tachibana S. Comparison of effects of root temperature on the growth and mineral nutrition of cucumber cultivars and figleaf gourd. Journal of Japan Society Horticulturae Sci., 1982, 51(3): 299-308.
80. Wilson C L, Solar J M, Ghaouth A El, etal. Rapid evaluation of plant extracts and essential oils from four aromatic wild plants in Israe. Journal of Chemical Ecology,1993, 19(6): 1229-1133.
81.YetisirH; Saris N. Effect of different rootstock on plant growth, yield and quality of watermelon. Australian journal of experimental agriculture, 2003, 42: 412-415.