鲜食葡萄组织培养快速繁育系统的建立

以鲜食葡萄品种无核白和美国早熟红无核（弗莱姆）为试材,利用当年生的半木质化嫩茎段作为组织培养的外植体,从外植体消毒、激素配比、培养基组分和移栽过渡等方面,对鲜食葡萄品种离体快繁系统进行了探索。结果表明：将当年生半木质化嫩茎段,先用75%乙醇消毒60 s、再用0.1%升汞（内加1%食盐）消毒8 min,获得葡萄单芽茎段无菌外植体;利用改良B5＋6-BA 0.5 mg/L＋IBA 0.1 mg/L培养基进行葡萄腋芽萌发启动诱导,产生初代苗;利用改良B5＋IBA 0.20 mg/L培养基作为继代生根培养基,将继代增殖培养与生根培养同时进行,能够减少愈伤组织过多的培养环节,提高基础瓶苗的使用率;在温度20～33℃、相对湿度≥65%条件下,光照强度从0.5万LX逐步上升到4.0万LX,光培炼苗4～7 d;待部分叶片长出瓶口形成角质层,有明显的反光感,茎秆充分转红后,选用粒径0.3～1.0 mm、pH值≤7.3的纯净河沙作基质进行沙培炼苗;当叶片明显增大,转绿,出现新生叶1～2片,叶表有光泽,长出白色侧根时,即可转入营养袋（或温室苗圃）炼苗;将驯化后的试管苗移栽到大田苗圃,只要营养袋土坨不散,一般成活率≥90%。将试管苗带叶接种扦插,对于加快繁殖速度、提高繁殖系数和节约时间具有重要意义。建立的该组培快繁系统,能够为建立优良品种的快速繁育体系提供有效可行的途径。

Tissue Culture Rapid Proliferation System Establishment of Table Grape

Taking ＇Thompson Seedless＇and＇American Early Red Seedless（Flem）＇ as tested grape materials,using half lignified tender stem segments as explants,tissue culture rapid proliferation system was explored and established,from the aspects of explant disinfection,phytohormones,medium components and transplanting transition.Proliferation steps were suggested：treating half lignified tender stem segments 60 s by 75% ethanol,then 8 min by 0.1% mercuric chloride（with 1% salt）to disinfect and get grapes single bud stem explants;using improved medium B5＋6-BA 0.5 mg/L＋IBA 0.1 mg/L to induce budding and produce early seedling;using improved B5＋IBA 0.20 mg/L as rooting medium and synchronizing multiplication culture and rooting culture,so as to reduce excessive cultivating steps and to improve basic plantlet usage;hardening the seedlings by light cultivation for 4-7 d under the condition of temperature 20-33 ℃,relative humidity above 65% and light intensity gradually increasing from 5 000 LX to 40 000 LX;when certain leaves growing out of bottle mouth,forming reflecting horny layer,and stalk turning enough red,hardening the seedlings by sand culture using pure river sand of diameter 0.3-1.0 mm,pH≤7.3 as substrate;when the size of leaf increasing obviously,turning green and 1-2 new leaves emerging,leaf surface showing luster and long white root growing out,hardening the seedlings by transferring them into nutrition bag（or greenhouse）;then transplanting domesticated test-tube seedlings to field nursery.As long as the earth lump in nutrition bag keeps tight,the survival rate could achieve more than 90%.Leaf inoculating the test-tube seedlings,could accelerate proliferation speed,enhance proliferation coefficient and save time.The tissue culture and rapid proliferation system provided an effective and feasible way for rapid proliferation of improved varieties.