蝴蝶兰NaN_3离体化学诱变及再生植株RAPD检测

采用NaN3对离体培养的蝴蝶兰类原球茎薄切片进行诱变处理,研究不同浓度和不同处理时间对类原球茎薄切片生长的影响,并对再生苗DNA进行了RAPD检测。结果表明,NaN3对类原球茎薄切片生长产生重要影响,部分薄切片白化或褐化死亡,再生类原球茎生长受到抑制,再生苗数量减少,部分再生苗畸变,叶片皱缩、变厚。10条RAPD引物的PCR检测发现,再生苗DNA序列发生了一定程度的变异,产生RAPD图谱带型多态性。根据以上结果认为,本体系合适的NaN3诱变剂量为6mmol/L,处理2d。     

马铃薯四倍体栽培种茎段组织的EMS诱变研究

用1%的EMS溶液处理“中薯2号”试管苗茎段材料,通过对诱变材料再生过程的观察、再生苗试管培养观察及棚栽阶段的生理指标统计分析,研究了EMS对马铃薯四倍体栽培种茎段组织化学诱变的作用。试验结果表明:EMS能够有效渗入生长点细胞;经EMS处理后试管苗生长量显著小于对照,再生植株和组织器官发生变异;试验证实1%(v∶v)浓度EMS处理4h以上具有明显的诱变效应,可以应用于今后马铃薯诱变育种。     

文心兰类原球茎的诱导与增殖技术的优化

以文心兰试管苗叶片为材料,在(25±1)℃,光照强度24～30μmol/m2.s,光照时间10 h/d下研究类原球茎的诱导和增殖技术。结果表明:①蔗糖适于作为类原球茎诱导的碳源,类原球茎诱导率52%,单位叶片类原球茎2.725个;②附加3～5 mg/L的Ad处理有利于类原球茎诱导(诱导率57%,单位叶片类原球茎数量3.725～4个);③6-BA是影响文心兰类原球茎增殖的主导因子,中等浓度的6-BA(3.0 mg/L)有利于类原球茎的增殖,增殖率(15.29)极显著高于其它水平;④培养基1/2MS+6-BA 3.0 mg/L+2,4-D 1.0 mg/L+NAA 0.4 mg/L+蔗糖20 g/L最有利于文心兰类原球茎的增殖,增殖率最高(15.95),极显著高于其它处理组合。     

化学诱变结合离体选择选育胡椒抗瘟病无性系

在胡椒(PipernigrumLinn.)茎尖丛生芽增殖技术的基础上,以印尼大叶种“LampongType”无菌实生苗作外植体源,采用EMS(ethylmethanesulphonate)对胡椒茎尖进行化学诱变处理,并结合辣椒疫霉(Phytophthoracapsici)培养滤液对胡椒茎尖及其增殖形成的丛生芽进行离体选择。结果表明,利用EMS在胡椒茎尖接种前进行化学诱变可提高抗瘟病变异体的产生频率。利用EMS不同浓度和不同时间对胡椒实生苗茎尖处理后接种,不经离体选择,直接在再生植株上进行抗病检测,抗性植株频率在4.3%￣26.67%之间,而不施用化学诱变的处理则未获得抗性植株。利用不同浓度EMS和不同诱变处理时间结合不同浓度的辣椒疫霉培养滤液离体选择,各种组合所获得的抗性植株比率在33.33%￣100%范围,但各种组合均较EMS和辣椒疫霉培养滤液单独处理的要高,其中以1.0%EMS处理60min结合25%辣椒疫霉培养滤液处理的最好。     

蝴蝶兰Co^60γ辐射诱变育种研究

以蝴蝶兰的原球茎为试验材料，设置ck、10GY、20GY、30GY、40GY、50GY等6个辐射剂量进行Co^60γ射线诱变处理。并对辐射后的原球茎进行分化扩繁生根落地栽培，观察田间生长变化，发现了诱变苗。     

铁皮石斛的组织培养与快速繁殖

以铁皮石斛(Dendrobium candidum Wallichex Lindley)无菌苗分成的茎段、带顶芽的茎段和根蔸3类外植体在MS＋6-BA0.5mg/L＋NAA0.2mg/L上均能诱导出丛芽,只有根蔸能诱导原球茎,且诱导丛芽倍数高于其它两类;在此培养基上原球茎的增殖系数高于其它组合的培养基;温度对原球茎继代增殖有明显影响,(25±1)益最适合原球茎的生长繁殖;温度对生根的影响较为明显,较低的温度(23℃)有利于减少分生苗的数量,形成健壮高大的试管苗,适量的香蕉能促进形成粗壮的根和苗。     

甜叶菊多倍体离体诱导技术体系的建立

以甜叶菊茎段为外植体,用不同浓度的秋水仙素在试管苗茎段再生植株过程中进行不同时间处理,探讨适合于甜叶菊离体多倍体诱导的最佳秋水仙素处理时间和浓度,初步建立甜叶菊离体诱导多倍体的技术体系。结果表明,秋水仙素对甜叶菊植株形成影响较大,秋水仙素处理的最佳剂量为0.1%处理3 d,再生植株总数高且多倍体比率达42.3%;同时,再生苗生根也受到了一定影响;多倍体苗叶片厚实、颜色较深、植株矮化,气孔增大,单位面积气孔数较少;多倍体细胞核体积增大,染色体数加倍。     

EMS处理对豇豆生长发育的影响

为了使EMS诱变在豇豆育种中发挥最佳效果,获得优质的诱变材料,对豇豆品种‘天畅9号’、‘银豇王’进行了3种EMS浓度、3种处理时间的诱导处理,并对当代的种子萌发、幼苗生长、植株主要农艺性状及生化指标的变化进行研究。结果表明:EMS能导致豇豆种子的发芽力和生长速率下降,叶片变小,而种子浸出液电导率、幼苗SOD活性及可溶性蛋白质含量增加;对银豇王的叶绿素含量、一级分枝数、结荚性有增加倾向,而天畅9号表现相反。处理剂量以浓度0.3%EMS、时间3~4 h为宜。     

春石斛离体培养条件优化及其试管开花研究

以春石斛类原球茎体及其试管苗为试验材料,采用L9(34)正交试验设计,比较不同激素浓度和不同浓度天然附加物对春石斛原球茎增殖和壮苗生根的影响,同时进行离体培养条件的优化,并在此基础上进行试管开花研究.结果表明:(1)最适宜春石斛原球茎增值的培养基为KC+1.5 mg/L 6-BA+0.5 mg/L IBA+100 mL/L椰汁;(2)最适宜春石斛试管苗壮苗生根的培养基为1/2MS+1.0 mg/L IBA+1.0 mg/L NAA+100 g/L马铃薯匀浆物;(3)在4种不同花色的春石斛中,玫瑰红花色的春石斛试管苗花芽诱导率最高,为38.67％;(4)磷含量增加至原来5倍时,春石斛试管苗花芽诱导率最高,为32.14％.     

夜间复合光质LED不同光照时间对水培切割再生生菜生长和品质的影响

以耐切割叶用莴苣品种‘奶油生菜’为试材,用LED光源对越夏水培切割再生生菜进行不同光照时长的夜间补光处理,研究暗期不同的补光时间(0、4、8、12 h)对水培生菜再生植株生长生理及营养品质的影响。结果表明:光照时间的延长,有利于加快生菜再生苗的地上部和地下部的生长速度,使再生生菜的可溶性糖含量显著增加,硝酸盐含量显著降低,有效地缓解高温季节再生苗受到的高温和切割伤害,显著提高了生菜再生苗的产量和品质,并且以延长光照时间为8~12 h效果更佳;其次,随着光照时间延长,再生生菜叶片光合色素的增加,叶绿素荧光主要指标有所提高,提高了叶片光合的同化能力,光抑制现象减弱,其中延长光照时间12h处理叶绿素含量显著或极显著高于其他处理,此外,适当延长光照时间,可增加生菜再生苗对矿质元素N、K、Ca的吸收。在本研究中,补光12 h处理整体最好,但是其产量品质与8 h处理差异不显著,结合生产实际和节能的角度综合考虑,对海南高温季节的水培切割再生生菜进行暗期LED补光,推荐适宜的补光时间为8 h。     

Studies on plant regeneration and somaclonal variation in Saintpaulia ionantha Wendl. (African violet)

Efficient plant regeneration of Saintpaulia ionantha (African violet) has been obtained in the present study. MS medium supplemented with 1.0 mg L(-1) IAA and 2.0 mg L(-1) Zeatin resulted in 100% shoot regeneration and induced the highest number of shoots (average 15.0 +/- 0.8 shoots per explant) after being cultured for 8 weeks. The above hormone combination was optimum for shoot regeneration. Most of Saintpaulia ionantha plantlets derived from tissue culture system could be hardened and transferred to the greenhouse conditions with 84.0 +/- 1.6% success rate. However, regenerated plantlets of Saintpaulia ionantha (even after 12-months-old) failed to flower. Morphological characters of regenerated plantlets of Saintpaulia ionantha were observed and compared with in vivo (intact) plants. Regenerated plantlets showed some differences in morphological characters, such as height and leaf size, texture and colour, but the plantlets showed no variation in leaf arrangement and leaf margin. However, the morphological characters of the regenerated plantlets were found to be unstable.     

甲基磺酸乙酯(EMS)对玉米自交系诱变效应的研究

用化学诱变剂甲基磺酸乙酯处理玉米自交系082的种子,研究了该诱变剂对玉米的诱变效应.结果表明:甲基磺酸乙酯对玉米自交系种子是一种有效的化学诱变剂,在M1代引起了较大的生理损伤和生物学效应,M2代质量性状未发生明显变异,但株高、穗位高、茎周长和穗上叶片长等数量性状出现了较高的突变频率,其中株高的负突变增加明显。     

蝴蝶兰叶片体细胞胚胎发生发育及其分化成苗的研究

蝴蝶兰幼叶离体培养直接诱导体细胞胚胎的发生并进一步发育成原球茎和分化成苗。体细胞胚胎发生起源于上表皮细胞或上表皮下方的叶肉细胞,为单细胞起源。单细胞原胚分裂形成多细胞原胚,历经球形胚、梨形胚、心形胚和子叶胚的发育过程,最终成为较大颗粒状的原球茎。较高浓度的苄基腺嘌呤(6-BA)和腺嘌呤硫酸盐(AdSO4)配合使用能有效诱导体细胞胚胎的发生,最高诱导率可达40%。适当降低6-BA和AdSO4浓度有利于原球茎分化成苗,但两者浓度过低苗的生长发育会受到影响。     

Haploid Plantlet Production through Somatic Embryogenesis in Anther-Derived Callus of Bupleurum falcatum

Abstract

This study was carried out to verify the production of haploid plantlets through somatic embryogenesis of Bupleurum falcatum in anther culture (2n=16). Flowers with anthers at the uninucleate stage, less than 200 µm in anther length, were exposed to 10ºC for 5 days (cold pretreatment) and the anthers were cultured on MS medium supplemented with 2,4-D and/or picloram at various concentrations at 30ºC. The optimal supplement for callus formation was a mixture of 0.075 mg L-¹ 2,4-D + 0.075 mg L-¹ picloram or 0.75 mg L-¹ 2,4-D without picloram. Only a few calli were induced from the anthers without cold pretreatment. The calli were transplanted to MS medium without phytohormones and cultured at 25ºC for plant regeneration. Among one hundred twenty root tips of the regenerated plantlets examined, 14.2% were haploid (n=8). However, in the plantlets regenerated from anthers without cold pre-treatment only 2.5% was haploid. In both haploid and diploid regenerated plantlets, the chromosome number was fixed without variation. Among the regenerated plantlets, one was albino. Haploid plantlets were transplanted to the field after acclimation in pots filled with vermiculite under 90% humidity for a month, and haploid plant were produced. The potential of haploid plants derived from anther culture for production of high-yield and good-quality cultivars is discussed.     

农杆菌介导的DnMADS2基因转化金钗石斛初步研究

为建立稳定有效的金钗石斛转化体系,进一步研究花发育相关基因的功能,为石斛兰分子育种提供基础,本研究以金钗石斛类原球茎（PLBs）为转化受体,通过农杆菌介导的方法将DnMADS2基因转入金钗石斛。结果表明：用20 mg/L潮霉素筛选4个月后获得抗性PLBs,对其进行HPT基因和DnMADS2基因的PCR检测表明,目的基因已成功整合到PLBs中。获得的转化PLBs在抗性培养基上分化为不定芽后,再经生根壮苗培养获得转化植株。对转化植株进行Southern检测,结果表明,DnMADS2基因已整合到5株金钗石斛转化植株基因组中。据此建立的稳定高效的金钗石斛转化体系和获得的转基因植株,将为DnMADS2基因功能的进一步研究和金钗石斛转基因育种奠定基础。     

Screening diploidSolarium genotypes responsive to different Anther culture conditions and ploidy assessment of Anther-derived roots and plantlets

Twenty-three diploid (2n=2x=24) potato clones were screened for response to four different anther culture media. Anther derived tissues (calli/embryos) were obtained from seven of the clones in liquid culture media. Anther culture response of these seven clones was further quantified on five additional media. Since clonal responses are specific to culture conditions (Tiainen, 1992), the seven genotypes somewhat differed from each other in the magnitude of the response to each of the media. In agreement with Heberle-Bors (1985) a more realistic assessment of the responsiveness was possible when pollen counts per anther were used to express the yield of anther-derived material. A relatively higher level of exogenous cytokinin (6-benzyladenine) to that of auxin (3-indoleacetic acid) proved to be favourable for the inducton of calli/embryos. Use of a semi-solid medium saved space and was convenient for culture observations therefore it was found more conducive for the genotypic screening. Anther derived tissues that regenerated to produce root tissue and/or plantlets were further multiplied on a root culture medium and a plantlet medium. Liquid culture conditions on orbital shakers were conducive for the growth of root explants. Anther derived material (roots and plantlets) originating from clone 9507–04 revealed differences in growth vigor. Flow cytometric estimation of ploidy in regenerated roots and plantlets, revealed that clone 9507–04 produced 44% monoploid roots and 77% monoploid plantlets. Seventy-three percent anther-derived plantlets from clone 6028–02 were monoploids.     

马铃薯叶盘愈伤组织再生苗抗青枯病变异株的筛选

摘要从340株马铃薯叶盘愈伤组织再生苗中经过试管苗接菌初筛选、温室抗性鉴定、人工病圃和田间自然病圃进一步鉴定获得了2株抗青枯病能力比母体显著提高的变异株.过氧化物酶同工酶分析表明,抗性变异株的酶活性都比母体高,其酶图谱比母体多了两条带,间接证明了它们在遗传上与母体有一定差异.     

马铃薯茎尖脱毒培养影响因素研究

以马铃薯茎尖为外植体,通过组织培养研究了不同的茎尖大小、不同浓度和配比的细胞分裂素和生长素对脱毒效果的影响。结果表明:茎尖越小,脱毒率越高但成活率和成苗率越差;茎尖越大,成活率和成苗率高但脱毒率差。6种培养基中以MS+NAA 0.1 mg.L-1效果最好,成苗率为33.75%,为本试验中茎尖诱导分化成苗的最优培养基。