青檀多倍体诱导试验初报

为获得具有优良性状的青檀四倍体植株,以青檀幼苗为试验材料,通过秋水仙素浸泡生长点的方法,研究了不同秋水仙素浓度和处理时间对青檀染色体加倍的诱导效果。结果表明：以0.4％—0.8％秋水仙素处理72ｈ诱导效果最佳,变异率最高达34.2％。经流式细胞仪和茎尖染色体鉴定,初次镜检青檀四倍体植株染色体数目为2n＝4x＝36,成功获得了青檀四倍体植株。     

白魔芋多倍体诱导研究初报

采用种子浸泡法和根状茎顶芽滴液法进行白魔芋多倍体诱导，比较了不同秋水仙素浓度和不同处理时间对出苗率和变异率的影响。结果表明，滴液法对材料的毒害性更小，经同一浓度秋水仙素溶液处理后出苗率和植株形态变异率均高于种子浸泡法，但诱变加倍的效果不如浸泡法。实验中获得一株纯合四倍体植株，由0.20%秋水仙素溶液处理种子48h诱变得到。     

秋水仙素诱导乌拉尔甘草同源四倍体的研究

在组织培养条件下,用秋水仙素溶液浸泡乌拉尔甘草试管苗(2n=2X=16)茎尖进行同源四倍体诱导,比较了不同秋水仙素浓度和不同处理时间对存活率、变异率的影响,并对加倍植株的鉴定和纯合四倍体的分离方法进行了研究.结果表明,秋水仙素浓度为0.1%,浸泡时间为12h时,茎尖的存活率和变异率分别为52.3%和22.1%,诱导率较高,并通过保卫细胞和根尖染色体数目鉴定,成功获得了甘草同源四倍体(2n=4X=32),为开展甘草多倍体育种奠定了物质和技术基础.     

芦笋单倍体染色体加倍技术研究

研究了芦笋单倍体染色体加倍的方法。在离体培养条件下,以秋水仙素为诱变剂,分别用浸泡法和培养基添加法处理芦笋单倍体幼苗的茎尖,比较了秋水仙素不同浓度、不同处理时间的诱导效果。结果表明,培养基添加法的诱导效果好于浸泡法,当在培养基中添加0.3%秋水仙素并处理7天时诱导效果最佳,染色体加倍频率与成活率分别可达82.50%和80%。加倍后的二倍体植株与单倍体植株相比,茎干变粗,气孔与保卫细胞增大,保卫细胞内的叶绿体数增多。     

秋水仙素诱导蚕豆及玉米根尖细胞染色体变异的研究

为进一步了解秋水仙素对农作物根尖细胞染色体畸变的遗传毒害效应,以地方品种敏感型绿皮小粒蚕豆和饲用玉米为材料,研究不同浓度秋水仙素（0、0.01%、0.05%、0.10%、0.15%、0.20%）及不同培养时间（24、48、72h）对2种作物染色体的畸变影响。结果表明：秋水仙素能诱发根尖膨大,当秋水仙素浓度为0.20%处理24h,蚕豆根尖膨大率100%;秋水仙素浓度为0.20%处理72h,玉米根尖膨大率65.8%;适当浓度秋水仙素（0.01%、0.05%）可促进蚕豆和玉米根尖细胞的有丝分裂,当浓度≥0.10%,反而抑制蚕豆和玉米细胞分裂。在染色体畸变类型中,微核最多,其次是染色体断片,最少是染色体桥。秋水仙素浓度0.20%,处理时间72h,蚕豆畸变率和玉米畸变率均最高,分别为11.92%和7.25%;秋水仙素浓度0.10%,处理时间72h,蚕豆染色体加倍指数最高为7.97%;秋水仙素浓度0.20%,处理时间24h,玉米细胞加倍指数最高为4.64%。     

秋水仙素浓度、浸苗时间对烟草单倍体幼苗的加倍效应研究

为了提高烟草单倍体植株的染色体加倍率,采用二次饱和D-最优设计,以两片真叶期的烟草单倍体幼苗为材料,研究了秋水仙素浓度及其浸苗处理时间对染色体加倍的效应.结果表明,随着秋水仙素浓度的提高,加倍率呈开口向下的抛物线曲线变化趋势;加倍率与浸苗时间呈线性回归关系,加倍率随浸苗时间的延长而增加;0.346 9%的秋水仙素、浸苗72 h是烟草单倍体幼苗染色体加倍率最高(达54.18%)的最佳处理组合.     

氨磺灵和秋水仙素诱导橡胶草四倍体效果的比较研究

为探讨不同诱变剂对橡胶草四倍体的诱导效果,以橡胶草幼芽为材料,采用浸芽法,比较分析氨磺灵和秋水仙素2种诱变剂不同浓度和处理时间对橡胶草多倍体的诱导效果及多倍体橡胶草的形态特征变化。结果表明：氨磺灵处理橡胶草的成苗率(60%~75%)和最高诱变率(14.08%)均高于秋水仙素处理（0~59%和5.56%）,其嵌合体率(27.27%)低于秋水仙素处理(75.00%),其中0.04 mmol/L氨磺灵处理 30 min,成苗率为71.00%,变异率14.08%,为最佳诱变条件。可见,氨磺灵比秋水仙素更适合用于橡胶草多倍体诱导。诱导后获得的四倍体橡胶草叶片厚度、气孔大小、花和种子大小均显著大于野生型二倍体植株,气孔密度则显著低于野生型植株,这些形态特征可辅助多倍体橡胶草快速筛选和鉴定。本研究初步探明了2种诱变剂对橡胶草染色体加倍的诱导效果,为今后橡胶草多倍体育种提供理论依据和技术支持。     

不同浓度秋水仙素对贡蕉的多倍体诱导

本试验以贡蕉胚性细胞悬浮系、胚性愈伤组织和多芽体为试验材料,利用不同浓度的秋水仙素溶液对不同外植体材料进行不同时间的诱变处理,以期获得染色体加倍的植株。结果表明,试验所用的胚性悬浮细胞系、胚性愈伤组织和多芽体三种不同外植体对秋水仙素处理的敏感性都较高,出现了不同程度的死亡,以0.4%(w/v)浓度的秋水仙素溶液处理多芽体48h诱导效果较好,多芽体的死亡率达到45.83%,接近半致死率,诱变率为18.64%。通过染色体计数和气孔鉴定,获得62株纯合四倍体小苗。诱变获得的植株的染色体数目加倍,叶片气孔数量减少,体积增大。本次试验为香蕉多倍体诱变育种和新种质的创新提供了技术参考和相关依据。     

利用抗微管除草剂胺磺灵诱导黄瓜四倍体

用Oryzalin对黄瓜茎尖进行四倍体诱导试验,结果表明,浓度为100~200μmol/L Oryzalin处理24 h对黄瓜茎尖离体四倍体诱导较为合适,可以有效代替秋水仙素。和秋水仙素相比,Oryzalin使用的浓度低,效果好,且毒性小,对人和环境比较安全,为诱导黄瓜染色体加倍的理想诱导剂。并用流式细胞仪对黄瓜离体诱导的幼苗进行了倍性分析试验。     

彩色马蹄莲2n配子育种技术初探

以彩色马蹄莲的栽培品种‘黒魔术’Black magic、‘火烈鸟’Flamingo为实验材料,研究彩色马蹄莲2n配子育种技术。选择刚从茎秆中抽出的幼小健壮小花,用0.15%的秋水仙素处理以诱导2n配子,将诱导的2n配子进行不同组合的杂交实验。结果表明：秋水仙素处理对彩色马蹄莲雄雌蕊发育有一定毒副作用,其中对雌蕊的损伤大于雄蕊;2n配子能在一定程度上克服杂交后代不育性;在相同条件下,经2n配子诱导处理的杂交种子与未处理的对照种子相比,种子萌发时间大幅度缩短（50天左右）,叶片变薄、变大、叶色变浅,有些杂交植株表现出不同程度的畸变。通过对杂交后代的细胞学鉴定表明：处理配子杂交后代中获得染色体数目为48（2n=3x=48）的三倍体植株。     

In vitro induction of tetraploids in Phlox subulata L.

Tetraploid plants of Phlox subulata L. were induced successfully by treating shoot tips in vitro with colchicine. Shoot tips excised from in vitro shoots were treated with four different concentrations of colchicine (0.005, 0.01, 0.02, 0.04%) in solid MS medium supplemented with 4.54 μM TDZ and 0.49 μM IBA for 10, 20 or 30 days, respectively. The survival rates of shoots tips were affected by the concentration of colchicine and the duration of treatment. High concentration and longer duration reduced survival of the shoot tips, but the effect of duration of colchicine was more than that of concentration. Tetraploid plants were obtained in all of the treatments, but the percentages of tetraploids varied among different treatments, from 25.0% to 75.0%. The most efficient condition for inducing tetraploids was to treat shoot tips with 0.005% colchicine for 20 days, with 30.0% survival rate of shoot tips and 6 tetraploid plants out of 10 plants examined. The rooted tetraploid plants were transplanted successfully in a solar greenhouse. Under the same growing condition, significant varieties in flower bud and flower sizes were detected between 2x and 4x plants. The flower diameters of tetraploid and diploid plants were 2.91 cm and 2.24 cm, respectively.     

Induction of <Emphasis Type="Italic">Anthurium andraeanum</Emphasis> “Arizona” tetraploid by colchicine in vitro

Tetraploids plants of Anthurium andraeanum “Arizona” were successfully induced after treating diploid tissue masses with colchicine. Masses originating from diploid aerial roots were treated with colchicine at three different concentrations (i.e., 0.1, 0.2, 0.3%) for about 3, 5 and 7 h, and then were transferred into Murashige and Skoog medium containing 3 mg/l BAP + 0.2 mg/l 2,4-D. After 60 days, the survival rate and numbers of regenerative shoots were scored. The high concentration and longer duration sharply reduced survival rate. In contrast, the regeneration of plantlets was not noticeably affected by colchicine. Tetraploid plants were obtained in all treatments, but the percentage of induced tetraploids ranged from 0.2 to 7.6%. The best induction was obtained with a 5-h, treatment with 0.3% colchicine. The stomatal size of tetraploid plants was larger than in diploid plants; however, the stomatal density was lower than in diploid plants. Tetraploid plants possessed stronger petioles, thicker and deeper green leaves, and thicker and longer lived spathes in comparison with diploid plants. Abnormal spathes, such as double spathes or those lacking pedicels, were observed in tetraploid plants. Tetraploid plantlets could be regenerated via aerial roots; this technique could be applied to tetraploid plant propagation.     

Short-duration colchicine treatment for in vitro chromosome doubling during ovule culture of Beta vulgaris L.

Colchicine uptake into ovules of sugar beet after 7 days of culture and its chromosome-doubling effect on ovule-derived plants were studied with high colchicine concentrations (0.4–6.0%) and short treatment duration (0–5 h). The best result of 4.2 diploid plants per 100 ovules was produced by treatment with 0.4% colchicine for 2.5 h. Both colchicine concentration and treatment time of ovules showed toxic effects on embryo formation, but it was stabilized at a low level with short exposure. The chromosome-doubling effect, by contrast, was unchanged with the colchicine concentrations used, but highly affected by the duration of exposure studied. A maximum percentage of 60% diploid plants was obtained after 3–5 h of uptake, which corresponds to only 31–39% of the total capacity for colchicine uptake in the ovules. Further uptake of the drug produced mainly toxic effects. Flow-cytometric measurements of the ploidy level in plantlets in vitro and of the same plants before flowering in soil were similar in about 80% of cases. Thus, flow-cytometric selection of diploid plants in vitro may be an efficient tool.     

Chromosome doubling of Allium fistulosum × A. cepa interspecific F1 hybrids through colchicine treatment of regenerating callus

Regenerating calluses of Allium fistulosum × A. cepa interspecific F₁ hybrids were treated in vitro with colchicine. A factorial experiment was designed to test the effects of colchicine concentration and time on the recovery of tetraploid plants from in vitro-colchicine-treated calluses. Shoot production of regenerating calluses following in vitro colchicine treatment decreased with increasing colchicine concentration and treatment time. Cytological analyses of root tip cells from regenerated plantlets showed that chromosomes of control plantlets (not treated with colchicine) were not doubled. Chromosome number of some plantlets regenerated from in vitro-colchicine-treated calluses were doubled, resulting in tetraploids. Calluses treated with 0.1 or 0.2% colchicine in BDS liquid medium for 48 or 72 hours yielded the highest numbers of tetraploid plantlets. Chromosome bridges at anaphase or early telophase were observed in diploid and tetraploid plants; their potential use is discussed. These results demonstrate that in vitro-colchicine treatment of regenerating calluses of interspecific F₁ hybrids is effective in recovering tetraploid plants. This revised version was published online in July 2006 with corrections to the Cover Date.     

濒危药用植物天山雪莲多倍体植株的化学诱导

以叶片诱导的愈伤组织为材料,用秋水仙素溶液浸泡法和秋水仙素加入培养基法分别进行诱导。实验结果表明,秋水仙素加入培养基法要好于秋水仙素溶液浸泡法,具体表现为:用秋水仙素加入培养基法诱导出的愈伤组织褐化死亡率低,分化率高,四倍体细胞平均比例高。而在处理浓度与时间的选择中,以高浓度的秋水仙素与短时间处理组合较好。秋水仙素的浓度为600 mg/L处理3 d效果最佳,四倍体细胞比例达41.5%。     

Efficient in vitro Chromosome Doubling During Beta vulgaris Ovule Culture

The effect of in vitro colchicine treatment of sugar beet ovules, after 7 days culture, on embryo formation, regeneration and ploidy of regenerated plants was studied with 5 concentrations of colchicine and 5 durations of treatment arranged as a 5 × 5 factorial in incomplete blocks. The best results were obtained with the shortest duration of treatment (5 hours) and the highest concentration of colchicine (0.4 %) giving 5.0 diploid plants per 100 ovules with 62.1 % of regenerated plants being diploid. Statistical analysis revealed that treatment effects could be separated into a toxic effect reducing embryo formation and a chromosome doubling effect affecting percentages of diploid regenerated plants. Toxic effects on embryo formation could be explained by simple exponential decay models, toxicity of the drug (decay constant) increasing linearly with duration of treatment. Duration of treatment had no effect on chromosome doubling percentages. The effects of colchicine concentration on chromosome doubling were explained by an exponential saturation model with spontaneous chromosome doubling of 8.1 % and saturation at 51.4 % diploid plants at 0.2 % and higher colchicine concentrations. In addition, treatments increased percentages of 4N and 6N plants from 0 % without colchicine to 10 % on average for treated ovules. A response surface model fitted to the total yield of diploid plants per ovule indicated that shorter durations of treatment and higher colchicine concentration may improve results.     

离心超重力诱发水稻变异效应初报

为了研究离心超重力是否对水稻产生诱变作用,通过采用9个不同的离心超重力和处理时间组合处理水稻品种矮秀占的萌动种子,并对处理后代进行研究。结果表明,随着处理的离心超重力加大和处理时间延长,发芽率下降,M2代的变异频率上升。处理效果以超重力为12000×g,处理时间60 min最好。处理M1代未发现明显的表型变异,M2代发生多样性变异,主要表现在茎秆、叶片、生育期、穗部性状、籽粒、稻米外观和直链淀粉含量,并可以稳定遗传。说明离心超重力不仅对水稻处理当代植株的生长发育产生抑制作用,而且可以诱发水稻产生明显的可遗传变异,可以作为一种新的诱变育种手段。