秋水仙素诱导中间锦鸡儿多倍体的研究

 以中间锦鸡儿（caragana intermedia）种子为试验材料,通过秋水仙素浸泡种子的方法研究了不同浓度及作用时间下秋水仙素对其染色体加倍的诱导效果。结果表明,直接浸泡种子,以0.3%秋水仙素浸泡48 h为最佳,诱变率为53.3%;种子经萌发12 h后再处理,以0.15%秋水仙素浸泡36 h诱变率最高,为73.3%。对变异株根尖进行染色体计数后发现,二倍体对照2n = 2x = 16,变异株2n = 3x = 24为三倍体,变异株2n = 4x = 32为四倍体,同时也发现存在嵌合体现象。     

春石斛的多倍体诱导

 在离体培养条件下比较了秋水仙素不同浓度、不同处理时间诱导杂种春石斛体细胞染色体加倍的效果。结果表明, 0.06%秋水仙素处理12 h的诱变效果最佳, 诱变率达69.1%。用组织培养结合不定芽诱导技术获得了稳定的多倍体变异株。变异株与正常株形态差异明显, 气孔数及大小之间差异显著。二倍体对照2n = 2x = 38, 变异株2n = 4x = 76 (四倍体) , 同时也发现存在嵌合体现象。     

秋水仙素诱导大葱多倍体的研究

 以章丘大葱种子为材料,研究了秋水仙素处理浓度和时间对大葱体细胞染色体加倍的效果,并利用形态观察,根尖细胞染色体计数及流式细胞仪等方法进行倍性鉴定。结果表明,0.8%秋水仙素处理48 h的诱变效果最佳。诱变植株存在明显“玻璃化”现象,生根困难,移栽后不能成活。通过组织培养去“玻璃化”,获得正常生长的四倍体植株。四倍体植株的细胞染色体数2n = 4x = 32,植株多倍体形态特征明显,植株生长健壮,叶色浓绿,叶面气孔变大,气孔密度减少;株高、葱白长度等与二倍体植株相比极显著增加。     

连翘二倍体与四倍体叶片特征比较

2009年采用秋水仙素溶液处理生长点的诱变方法和茎尖细胞染色体计数检测及流式细胞仪倍性鉴定方法首次诱导出连翘多倍体.经茎尖细胞染色体计数检测,变异植株茎尖细胞染色体数目为2n=6条,而原二倍体的染色体数目为2n= 28条,因此,变异植株(2n= 56)为四倍体.经流式细胞仪倍性分析验证,连翘四倍体体细胞DNA相对含量是...     

秋水仙素诱导三种紫薇多倍体的研究

 研究了秋水仙素处理浓度和时间对3种紫薇幼苗染色体加倍的诱导效应，通过染色体计数、流式细胞仪以及形态观察等方法进行了倍性鉴定。结果表明：秋水仙素诱导3种紫薇均得到了四倍体,0.5%～0.8%的秋水仙素处理紫薇和银薇48～96 h的变异率较高，其中以0.5%秋水仙素处理紫薇72 h的变异率最高，达54.17%。根尖染色体压片表明四倍体染色体数为：2n=4x=96，二倍体对照为：2n=2x=48。四倍体表现出叶片长、宽、厚均增大，气孔密度减小，气孔保卫细胞增大，保卫细胞内叶绿体数增加，花粉粒直径增大，花径、花瓣长、花瓣基部爪长均较二倍体明显增加，单朵花观赏价值大大提高。     

彩色马蹄莲多倍体的诱导

 在离体培养条件下,比较了不同浓度、不同处理时间的秋水仙素诱导彩色马蹄莲体细胞染色体加倍的效果。经形态学和气孔观察,0.05%的秋水仙素处理24 h的诱变效果最佳,诱变率高达52%。用组织培养结合不定芽技术对变异株进行稳定,获得了稳定的多倍体。对变异株进行细胞学研究后发现,二倍体对照2n=2X=32,变异株2n=4X=64,为四倍体,另外还发现六倍体细胞的存在。     

姜四倍体离体诱导及其形态学分析

以丛生芽为材料,在离体培养条件下比较了秋水仙素不同浓度、不同处理时间诱导姜体细胞染色体加倍的效果。结果表明:以0.20%秋水仙素液体培养基处理8d的诱导效果最好,诱变率和成活率分别达到了42.86%和70.00%。根尖染色体压片检查发现,四倍体染色体数为2n=4x=44,二倍体对照为2n=2x=22。四倍体与二倍体相比表现出植株高大,茎秆变粗,叶片长、宽、厚均增大,叶片下表皮气孔密度减小,气孔和保卫细胞增大,保卫细胞内叶绿体数增加等特征,其中气孔密度及保卫细胞内的叶绿体数可作为鉴别四倍体与二倍体的重要性状。     

君子兰未成熟胚四倍体诱导及染色体数鉴定

 以君子兰（Clivia miniata Regel.）品种‘胜利’为试材,采用一种新型外植体未成熟胚,利用秋水仙素与组织培养相结合的方法,研究秋水仙素浓度、处理时间和不同胚龄对未成熟胚诱导加倍的影响。结果表明：在MS + NAA 2.0 mg · L-1 + BA 1.5 mg · L-1培养基上,0.01%秋水仙素处理180 d胚龄的未成熟胚20 d诱变效果最佳,四倍体诱变率最高,达30.8%,形态变化率达36.5%,死亡率为34.1%。经0.03%秋水仙素处理120 d胚龄的未成熟胚30 d诱变效果也较好,四倍体诱变率较高,达24.0%,形态变化率达21.1%,死亡率为46.7%。经染色体数观察,加倍处理后代中存在2x、4x和混倍体植株。随机调查了267株加倍处理植株,得到30株四倍体（2n = 4x = 44）、194株二倍体（2n = 2x = 22）和43株混倍体,平均四倍体诱变率为11.2%。秋水仙素处理出现的植株生长缓慢,生根明显较晚,叶片变宽变短等形态变化可以作为加倍的初期有效判断指标。     

切花非洲菊多倍体诱变初报

 以非洲菊‘阳光海岸’和‘白马王子’ (2n=2x=50) 品种为供试材料, 采用秋水仙素对其组培丛生芽进行离体诱导, 成功获得了四倍体植株。秋水仙素处理浓度和处理时间分别为0.02%、0.05%、0.10%和24、36、48 h。试验结果表明, 以0.10%的秋水仙素处理48 h诱导效果最佳, ‘阳光海岸’和‘白马王子’的丛生芽变异率分别为16%和10%。总体上看, 非洲菊对秋水仙素的敏感性较低。经气孔鉴定、染色体计数和形态学观察, 变异材料染色体数加倍(2n=4x=100) 。与二倍体相比, 变异材料气孔面积增大, 花序变大, 叶色变深, 叶尖变钝圆, 叶片质感变厚, 花梗直径变粗, 花梗基部花青素着色变深。最终分别获得稳定的‘阳光海岸’和‘白马王子’多倍体植株200余株和300余株。     

豇豆四倍体的诱导及鉴定

用浓度为0.1%~0.3%的秋水仙素处理二倍体豇豆子叶期的茎尖生长点,获得同源四倍体(2n=4x=44),并对其进行形态学、细胞学、农艺学鉴定。结果表明:经0.2%秋水仙素处理4次获得同源四倍体的效果最佳,四倍体诱变率为2.809%;与二倍体相比,四倍体植株叶片、花器官、荚果等均表现巨大性,气孔密度和单个荚果种子数降低;用流式细胞仪进行倍性鉴定,豇豆二倍体DNA相对含量为250,四倍体为500,与根尖染色体鉴定结果一致。     

杜仲四倍体与二倍体光合特性比较

采用Li-6400便携式光合测定系统对杜仲四倍体和二倍体植株进行光合速率日变化、季节变化、光响应曲线和CO2响应曲线测定。结果表明:杜仲四倍体和二倍体净光合速率的日变化呈双峰曲线,中午存在明显的光合"午休"现象。杜仲四倍体的全天最大净光合速率显著高于二倍体。四倍体与二倍体的光合速率季节变化呈双峰曲线,峰值分别出现在6月和8月。四倍体与二倍体的LCP在18.00～27.78μmol.m-2.s-1之间,LSP在1 216.67～1 240.00μmol.m-2.s-1之间。CCP在72.50～117.75μmol/mol,CSP在1 347.47～1 397.48μmol/mol之间。杜仲四倍体的光合性能优于二倍体。     

In vitro tetraploid induction and generation of tetraploids from mixoploids in Astragalus membranaceus

This paper describes an efficient colchicine-mediated technique for the in vitro induction of tetraploids in Astragalus membranaceus and its confirmation by flow cytometry. Buds immersed in 0.2% colchicine solution for 36 h prior to culture induced as high as 35.3% tetraploid plants. Colchicine-induced tetraploids remained stable after 6 months in soil. Leaf characteristics of diploids and tetraploids in A. membranaceus were compared. It was determined that leaf sizes of glasshouse-grown plants and stomatal sizes of both in vitro and glasshouse-grown plants were suitable parameters for identifying putative tetraploids in A. membranaceus. As well as generating tetraploids, this technique generated mixoploids in A. membranaceus. Calli derived from mixoploid leaves, were induced to form buds and shoots. Individual shoots were classed as diploid, mixoploid and tetraploid by flow cytometry. This callus-based technique can be employed when a genome-doubling agent generates mixoploids but no tetraploid.     

杜仲大树移植与管理技术

杜仲是我国特有的经济树种,属国家二级保护中药材.因其大树移栽成活率较低,对杜仲大树移植过程进行了详细地阐述,提出移植过程及栽后管理中应注意的技术环节和应采取的技术措施.     

金冠苹果成熟胚离体诱导四倍体的研究

 以自然授粉的‘金冠’苹果种子为材料，研究了秋水仙素处理成熟胚离体诱导四倍体技术和嵌合体分离方法。结果表明：将完整胚在0.5%秋水仙素+1%二甲基亚砜的混合溶液中暗处理48 h，然后接种在MS+TDZ 1.0 mg·L^-1+IBA 0.5 mg·L^-1+蔗糖30 g·L^-1+琼脂6 g·L^-1的再生培养基中，变异率高达10.6%。与二倍体相比，四倍体试管苗生长缓慢，节间变短，叶片增大，叶形指数趋小化，气孔密度降低，保卫细胞增大。经压片法鉴定茎尖染色体数和流式细胞仪分析DNA含量，表明多数变异植株为同质四倍体。利用叶片离体再生器官技术，从诱导获得的嵌合体植株中分离出同质突变体。     

番木瓜四倍体的诱导及形态学分析

在离体培养条件下,将二倍体穗中红番木瓜无菌试管苗茎尖浸入0.05%、0.1%、0.2%、0.4%秋水仙素水溶液中处理1、2、3、4、5d,诱导多倍体,对照用无菌水处理2d。结果表明0.2%的秋水仙素溶液浸泡处理3d效果最好,诱导率为36.7%。通过体细胞染色体数目检测,变异芽为四倍体或嵌合体,四倍体染色体数目为36(2n=4x=36)。对获得的四倍体和二倍体进行了形态及气孔特征的比较,结果表明四倍体与二倍体植株在叶长、叶宽、叶厚、气孔大小、气孔密度、气孔保卫细胞大小及叶绿体数目等方面存在明显的差异,其中叶片厚度、气孔密度和保卫细胞内叶绿体数目可作为鉴别四倍体与二倍体的重要性状。     

Morphological and cytological studies of diploid and colchicine-induced tetraploid lines of crape myrtle (Lagerstroemia indica L.)

The tips of cotyledon-stage seedlings of three crape myrtle cultivars (“Zi Wei”, “Hong Wei” and “Yin Wei”) were treated with colchicine. Various concentrations of colchicine and different treatment durations were tested. Seedlings of “Zi Wei” treated with 0.5% colchicine for 72 h and seedlings of “Yin Wei” treated with colchicine (0.2% for 96 h, 0.5% for 48 h and 0.8% for 72 h) demonstrated high rates of mutation; “Hong Wei” showed a slightly lower rate. The highest rate of morphological variation was 54.17% and this was achieved when tips were treated with 0.5% colchicine for 72 h. Putative tetraploid plants were identified with morphological and cytological variations, such as larger and thicker leaves, darker green coloration, larger stomata, lower density of stomata across the lower leaf epidermis and increased numbers of chloroplasts per stomata guard cell. Flow cytometric analysis demonstrated that the nuclear DNA content of 15 of these putative tetraploids (of which 7 were “Zi Wei”, 5 were “Yin Wei”, and 3 were “Hong Wei”) was indeed doubled relative to that of control diploid plants. The chromosome number of the tetraploid plants was 2n = 4x = 96, while that of the control diploid plants was 2n = 2x = 48. Ornamental characteristics were also enhanced in the tetraploid plants, with increased diameter of the individual flowers and greater basal length of the petals and claws. Pollen diameter and the size of capsules and seeds were also significantly greater than those of diploid plants.     

生长素对杜仲愈伤组织诱导及增殖的影响

以杜仲幼茎为外植体,采用正交实验设计,添加不同种类及不同浓度的6-BA和NAA,组成15种不同的愈伤组织诱导培养基,对杜仲愈伤组织进行诱导增殖培养。结果表明:诱导培养基中添加1mg/L 6-BA+0.1mg/L NAA时,有利于杜仲愈伤组织形成,诱导率达86.67%;增殖培养基中添加0.5mg/L 6-BA+0.5mg/L NAA+4%蔗糖,能明显提高杜仲愈伤组织增殖系数。     

DNA ploidy level of colchicine-treated hops (Humulus lupulus L.)

The genome doubling agent colchicine has been used effectively to obtain tetraploid plants when starting with diploid material. Since the plant meristem consists of many cells, as well as obtaining tetraploids, it is possible to obtain mixoploids (chimeras consisting of diploid and tetraploid tissue) after colchicine treatment. Flow cytometry was used to determine the DNA ploidy level of different tissues of individual hop plants following colchicine treatment of diploid hops. Plants that are shown to be mixoploid after analysis of leaf tissue can display higher levels of tetraploid nuclei in root tissue, such that they could be mistakenly classified as tetraploid. The hop variety Galena-4n, which was previously considered to be a tetraploid, is shown to be a mixoploid based on leaf material. However, Galena-4n consistently produces triploid progeny when crossed with a diploid hop variety, indicating that Galena-4n produces tetraploid reproductive tissues and is effectively a tetraploid from a breeding perspective. The results from this study indicate that care should be taken when determining the DNA ploidy level of an individual plant after exposure to genome-doubling agents based on single tissues and that leaf tissue is a choice material for flow cytometric analysis.     

利用二硝基苯胺类除草剂离体诱导西瓜四倍体

 以二倍体西瓜单系Z-A和Z-R为试验材料,采用不同浓度的二硝基苯胺类除草剂——胺磺灵、二甲戊乐灵和氟乐灵处理其离体子叶,并以秋水仙素和不含任何诱导剂的液体培养基为对照,研究诱导西瓜四倍体的效果。结果表明,3种除草剂诱导西瓜四倍体的效果都比秋水仙素好,二甲戊乐灵、胺磺灵、氟乐灵和秋水仙素的四倍体诱导率分别为40.0%、33.3%、26.7%和13.3%。采用子叶直接处理比间接处理的四倍体诱导率高。通过 FDA荧光法对西瓜试管苗叶片表皮保卫细胞叶绿体计数,进行四倍体变异早期鉴定,准确率为90%以上。