【作者】 王素君；

【导师】 张毅功；

【作者基本信息】 河北农业大学 ， 植物营养学， 2004， 硕士

【摘要】 本试验研究目的是探索花叶爬山虎和五叶爬山虎离体快繁的有效途径，采用带腋芽茎段为主要外植体来诱导增殖。同时还分别利用花叶爬山虎和五叶爬山虎的叶片、茎段和叶柄来诱导愈伤，通过分化来增殖。试验以单因素和复因素设计，用方差分析和新复极差多重比较进行数据统计分析。着重研究并筛选爬山虎组织培养的最佳外植体，培养程序及培养基配方，主要结果如下： 1．花叶爬山虎和五叶爬山虎最佳的采样时期都为4月和7月，萌芽率高，无菌苗生长快；外植体以项芽以下3～7节位上的饱满腋芽为最好，污染率低，萌芽率高，成活后幼苗生长健壮。 2．花叶爬山虎诱导愈伤最适的外植体是叶柄，主要诱导激素为NAA，培养基组成：1／2 MS+BA 0.5 mg／L+NAA 0.1 mg／L+PVP 300 mg／L+白砂糖30g／L+琼脂7.5g／L，出愈早，量多，绿色，有生活力；五叶爬山虎诱导愈伤最适的外植体是茎段，培养基成分：1／2 MS+BA 0.5 mg／L+2，4-D 0.8 mg／L+白砂糖30 g／L+琼脂7.5g／L，愈伤浅绿色，质地疏松，有生活力。 3．继代培养过程中，花叶爬山虎易褐化，附加PVP 300 mg／L可减轻褐化程度，花叶爬山虎较适宜生长的基本培养基为GS和1／2MS，同时附加0.06 mg／L的NAA和0.5 mg／L的BA；而五叶爬山虎则在以1／2 MS为基本培养基中生长较好。 4．五叶爬山虎植物激素的选择，在NAA、IBA和IAA三种生长素中，以NAA为最适宜，增殖阶段需要浓度一般较低，0.02 mg／L～0.08 mg／L，配合一定量的细胞分裂素，就能促进无菌苗茎的伸长；诱导增殖的细胞分裂素，以BA为最好，浓度在1.5 mg／L时，配合0.02 mg／L～0.08 mg／L的NAA，30天的最高增殖系数一般可达到3.5；KT诱导增殖的能力较弱，低浓度的条件下可诱导五叶爬山虎生根。 5．爬山虎对于pH的适应范围很广，在pH为4.6～6.4的环境中都能生长，最适宜的pH范围是5.4～6.2，无菌苗生长健壮，茎伸长量大；基本培养基中铵态氮和硝态氮浓度的高低也影响五叶爬山虎的茎伸长量和增殖，过高或过低都不能使五叶爬山虎达到最佳生长状态。 6．以植物激素和有机添加物的正交实验，综合多因素影响，适合五叶爬山虎增殖快繁的最佳培养：1／2 MS+NAA 0.04 mg／L+BA 1.0 mg／L+KT 2.0 mg／L+LH 100mg／L+白砂糖30g／L+琼脂7.5g／L；1／2 MS+NAA 0.08 mg／L+BA 1.5 mg／L+KT 1.0 mg／L+LH 100 mg／L+白砂糖30g／L+琼脂7.5g／L，植株生长健壮，培养30天，增殖系数一般可达到3，若接种母体生长粗壮，增殖系数可达更高。 7．在生根培养中，最佳的生根培养基：1／2 MS+IAA 0.5 mg／L+IBA 0.3 mg／L，1／2 MS+NAA 0.06 mg／L+IBA 0.5 mg／L，分别添加白砂糖20g／L和琼脂7.5g／L，生根率100％，平均根数在3.5以上，接种的无菌苗一般在2.0cm左右，带两片叶片，长势好的材料生根效果更好。 8．炼苗移栽阶段，无菌苗的较适宜炼苗天数为10天左右，30天后成活率达到80%，锻炼过程适当延长炼苗天数能提高移栽成活率。移栽基质采用蛙石或者蛙石与沙各半为适宜，30天后成活率达到80%以上，而蛙石与营养土各半的配比，在生长后期，能为幼苗提供丰富的营养元素，使移栽苗生长健壮。 9.光合的测定，五叶爬山虎有明显的“午休”现象，13二00净光合速率最低，下午16:00有一高峰，以后净光合速率骤减。更多还原

【Abstract】 The stem of Silvervein Creeper and Virginia Creeper was used as explant to establish the rapidly multiplication technique systm of tissue culture, In addition still utilized the leaf and stalk passage and the petiole of Silvervein Creeper and Virginia Creeper to induction callus and multiplication .With single factor and orthogonal design and analysis of variance, the study aims to find out the optimum explant and best medium composition and the culture process in Silvervein Creeper and Virginia Creeper. The results were as follows:1 The optimum sampling period of Silvervein Creeper and Virginia Creeper is April and July, the germinating rate is high, and grows quickly; The axillary bud on the length location of the body with the below 3-7 of terminal bud is best explant, and pollution frequency is low, the germinateing rate is high, and the seedling grows robustly after survival.2 The optimum explant of Silvervein Creeper to adduction callus is the petiole, the main adduction hormone is NAA ,culture medium : 1/2 MS + NAA 0.1 mg/L + BA 0.5 mg/L + white sugar 30 g/L + agar 7.5g/L, adduced callus by it more early, quantity far more , the greener, differentiatia frequency higher; The optimum explant of Virginia Creeper to adduce callus is the stalk passage, the culture medium constitutes: 1/2 MS + 2,4-D 0.8 mg/L + BA 0.5 mg/L + white sugar 30 g/L + agar 7.5 g/L, the callus is light green, the quality of the callus is loose , differentiatia frequency higher .3 In differentiatia and subculture stage, Silvervein Creeper easy brown, and attachs PVP 300 mg/L alleviate the brownization degree. The fundamental culture medium of more fit growth of Silvervein Creeper is 1/2MS and GS, that attachs BA 0.5 mg/L, and NAA 0.06 mg/L; but Virginia Creeper are living in takes 1/2 MS as the fundamental culture medium to grow better.4 The optimum plant hormone to Virginia Creeper is chooseed, three kinds of growth hormones is NAA, IBA and IAA, and NAA is the most suitable hormone, the lower concentration of it in reproduces stage, NAA 0.02 mg/L~0.08 mg/L is connected with the cytokinin which may accelerate seeding growth; The cytokinin that the adduced multiplication is BA act as goodly, coordinates the NAA 0.02 mg/L~0.08 mg/L the multiplication coefficient attainability up 3.5 when the concentration is 1.5 mg/L, The capability that KT’s guidance is reproduce is infirm, but below the concentration KT may lead rooting.5 The Virginia Creeper is very vast as to the accommodation of pH, and is living in the environment of pH 4.6-6.4 wholly to grow, the most fit limit is pH 5.4-6.2 where the seeding grows robustly, and stalk extending amount is greatly; Ammonium nitrogen and the saltpetre nitrogen concentration height also affects stalk extending amount and multiplication coefficient of Virginia Creeper in the fundamental culture medium, and overtops either overlow are all not to be fit for.6 With the plant hormone is connected with organic accretion orthogonal design test, being fit for Virginia Creeper reproduces quickly optimum culture medium: 1/2 MS +NAA 0.04 mg/L + BA 1.0 mg/L + KT 2.0 mg/L + LH 100 mg/L + white sugar 30 g/L + agar 7.5 g/L; 1/2 MS + NAA 0.08 mg/L + BA 1.5 mg/L + KT 1.0 mg/L + LH 100 mg/L + white sugar 30 g/L + agar 7.5 g/L, after culture 30 days, the multiplication coefficient attainability up 3 and grows robustly, it is taller to reproduce the factor attainability.7 In adduction rooting stage, the optimum culture medium adduction rooting: 1/2 MS + IAA 0.5 mg/L + IBA 0.3 mg/L, 1/2 MS + NAA 0.06 mg/L + IBA 0.5 mg/L, at the same time attachs agar 7.5 g/L and sugar 20 g/L the rate of indduction rooting leading to 100%, the seeding high 2.0 cm, brings two leaves blade, the better stuff and the better redduction rooting.8 Acclimatization and transplants, the more suitable acclimatization time is 10 days, and the survival rate after 30 days attains 80% and the transplanting survival rate may more high with prolongs the acclimatization time.Transplanting basic matter adopts vermiculite or ve更多还原