【作者】 李江；

【导师】 孙仲序；

【作者基本信息】 山东农业大学 ， 果树， 2004， 硕士

【摘要】 本试验于 2002~2004 年在山东农业大学和烟台龙口复发中记公司进行。针对当前植物组培过程中出现的程序烦琐、培养成本高、移栽成活率低等问题,以大樱桃优质矮化砧木吉塞拉(Gisela)作为试材,对植物组培技术体系进行了系统的研究,从而对整个组培过程进行优化,得出较好的培养效果和大批量的优质苗木;主要结果如下:1. 在前人研究的基础上进一步进行研究,获得高效优良的无菌外植体。具体结果如下:(1)取材季节选在春季,室温条件下培养的植物材料各项指标要好于光照培养箱中培养的植物材料。(2)除已知的培养材料自身的影响外,培养基中的无机盐增加了外植体的褐化率;而加入活性炭等吸附剂可以减轻褐化率。(3)不同的灭菌方法研究结果表明:升汞加升汞的灭菌方法较为合适,高效优质无菌外植体的获得率相对较高。2.水的替代。 采用不同地区的自来水,用蒸馏水作对照配制培养基研究其对分化率和生根率的影响。研究结果表明:在生产中可以采用优质的自来水代替蒸馏水,简化了程序,降低了成本。3.分化条件的优化。 在分化培养基中,较佳的条件为: 7g/l 的琼脂,5.0～6.0 之间的pH,处于 1500～4000lux 之间的光照强度, 23~25℃之间的室温条件,30g/l 的糖;在采用较佳用量的条件下,采用 6-BA 的分化效果要大大好于 KT 的分化效果,且继代分化的效果仍然较好;以糖、6-BA、IBA 的较佳用量设计正交试验(L9(33)),根据正交表的结果得出三者的最佳用量为 MS+糖 30g/l +IBA0.3mg/l +6-BA0.8mg/l,并分析得出分化中的最大影响因素为 6-BA;每瓶培养基的经济用量为 35ml;在生根前期,将原有分化配方中激素的用量成比例缩小 2 倍,茎伸长效果较佳;且可以因此提高驯化成活率,提高质量。 1<WP=9>李江:吉塞拉大批量组培快繁条件优化的研究4.培养基中各营养成分的作用(1)大量元素在分化中的作用:大量元素主要起供应组培苗生长所需要的基本营养成分的作用。(2)微量元素在分化中的作用:微量元素在组培苗的健壮性方面作用较大,而对增殖系数的影响不大。(3)有机物在分化中的作用:有机物在组培苗的生长状况方面影响较大,而对增值系数的影响不大。(4)铁盐在分化中的作用:铁盐在组培苗的新生叶片及分化芽的颜色影响较大,缺失的情况下,叶片变黄;但对分化率影响不大。(5)激素对组培苗的分化至关重要。5.再利用的研究。 培养基再利用的较佳用量为重新加入 10%的原有琼脂用量、40%的原有 MS培养基营养成分和原有分化配方 100%的激素用量,结果能够达到原有的分化效果,且生长较好。6.瓶内生根条件的优化 在生根过程中,较佳的生根条件为:6g/l 的琼脂, 5.0~6.0 之间的pH,20g/l 的糖;将 IBA 、IAA、NAA 的较佳用量进行正交试验(L9(33)) , 得 出 较 佳 的 生 根 配 方 为 MS + IBA 0.3mg/l+ IAA0.3mg/l +NAA0.3mg/l,其最大的影响因子为 IBA;接种生根培养基后,可使之处于温室自然光条件下生根,但须控制光强在 1500～4000lux 之间,这样可以节省电能,有效地利用组培室,降低成本;每瓶生根培养基较佳用量为 30ml。7.瓶外生根条件的研究 瓶外生根基质以沙和草炭土为 1:1 的条件生根效果较好,驯化条件以盖封口膜保持湿度的方法较好,并将光强保持在 1500～4000lux 之间;生根效果最好。8.体系的优化 2<WP=10>山东农业大学硕士学位论文综合以上的优化结果,分析得出较佳的一套合理可行的批量生产大樱桃砧木吉塞拉的程序,其成本仅是常规育苗成本的 26.1% ,且生产的苗木品质优异,数目巨大,时间短,适合大规模的工厂化育苗更多还原

【Abstract】 The test has been done in Shandong Agriculture University and FHTKCompany of Longkou Yantai from 2002 to 2004. Because of the question ofcomplicated process, high cost and low living ratio, the system of plant tissueculture has been done when cherry’s avail named Gisela as material. Based onthe result, the total process of tissue culture in Gisela has been optimized, theresult is very good and a lot of seedlings which has good quality has beenowned, It is:(1) After choosing plant material in spring, the better culture condition ofplant material should be set under natural temperature, not in illuminatingincubator, then we will get high living rate asepsis explant.(2) Besides the effect of the culture material, more mineral element will addthe plant’s browness and the sorbent like active carbon will reduce browness.(3) Among different sterilization methods, the better method should be Hg+Hg, if we do this, the good asepsis explant can be got at high percent.2. Substitute of water Tap water from different area was made culture medium in order to studythe effect on multiplication process and rooting when distilled water as control,tap water which has high quality can replace the disitilled water and reduce thecost, optimizate the process.3.The optimization of multiplication In Multiplication process of Gisela’s tissue culture, the quantity of agarshould adopt 7g/l, pH should be set on from 5.0 to 6.0, light intensity shouldbe placed from 1500 to 4000lux, sugar is 30g/l, room temperature should becontrolled from 23 to 25, hormone should adopt 6-BA, 6-BA has moremultiplication than KT and still lies in high multiplication. The test oforthogonal test has been done based on the three factors of sugar, 6-BA, IBA,the best quantity should be adopt such quantity: agar is 7g/l, IBA is 0.3mg/l, 6- 4<WP=12>山东农业大学硕士学位论文BA is 0.8mg/l , and sugar is 30g/l, among this three factors 6-BA has thebiggest impact on the multiplication process. Before rooting process, let thequantity of all hormones change at the same time to 1/2 of original quantity,the microcutting will turn into the better one which is taller and more strong,the seedlings of high quality can be got at large number.4. Effect of nourishment of Ms culture medium(1) Macroelement’s effect: Macroelement in plant tissue culture has theefffect of giving the nourishment of the microcutting in plant tisue culture.(2) Microelement’s effect: Microelement in plant tissue culture has the efffectof giving the haleness of the microcutting in plant tisue culture.(3) Organic compound ‘s effect: Organic compound in plant tissue culture hasthe efffect on the growth condition, while has little effect on multiplication.(4) Ferric components’s effect: Ferric components in plant tissue culture hasgreat effect on the new leaves and new buds, If lacked, new buds and newleaves turn yellow, but multiplication don’t change when compared withnormal.(5) Hormone is very importantin to the process of the multiplication ofmicrocutting in plant tissue culture.5.The study of second use of culture medium The study tell us that adding 10% sugar, 40% nourishment of MSculture medium and 100% hormones of original quantity to the used culturemedium, the best multiplication can be get to the new culture medium, andgrowth condition of Gisela’s tissue culture is good.6. Optimization of rooting in vivo In rooting process, the better quantity is follow: agar is 6g/l, sugar is20g/l, pH should be placed from 5.0 to 6.0, In the three factors of IBA ,IAA, NAA,the better culture medium should be 6 g/l agar and Ms culturemedium, IBA which is 0.3, IAA which is 0.3, NAA which is 0.3 , the biggestfactor is IBA in rooting culture medium for Gisela, After playingmicrocuttings at bottle which aimed at rooting , then the culture of rooting canbe placed at nature light of greenhouse (nature light should be controlled from1500lux to 4000lux更多还原