秋水仙素诱导青岛百合四倍体研究

以青岛百合试管苗为试验材料,采用浸泡法和混培法研究了秋水仙素对青岛百合进行染色体加倍的诱导效果。结果表明:以0.05 %的秋水仙素浸泡处理24h的诱导效果最佳,诱导频率高达53.3%;经秋水仙素诱导的多倍体与正常二倍体植株比较,植株叶片变大,根系粗壮,气孔显著增大而单位面积气孔数减少。对变异材料的细胞学研究发现, 四倍体植株的染色体数目为2n=4x=48。     

迎阳报春四倍体诱导及鉴定

在离体培养条件下,研究了秋水仙素溶液处理对迎阳报春愈伤组织分化和多倍体诱导的效应。结果表明,秋水仙素处理明显抑制愈伤组织的分化,使死亡率增加;在试验浓度和时间范围内,以30mg·L-1秋水仙素溶液处理愈伤组织10d,四倍体诱导率最高,为23.08%;四倍体植株与二倍体植株相比,其气孔变大,且叶片变厚、变大,叶色浓绿、株型紧凑;二倍体植株染色体2n=2x=24,四倍体植株染色体2n=4x=48。     

秋水仙素诱导甘菊多倍体研究

以甘菊种子为试验材料,研究了不同秋水仙素浓度和处理时间对甘菊多倍体诱导的影响。结果表明:500mg/L的秋水仙素水溶液浸渍种子12h,变异率为23%,达到了极显著差异。根尖染色体压片检查表明,四倍体染色体数为2n=4x=36,并且获得了同源四倍体甘菊1株(2n=4x=36)。四倍体植株表现出叶片下表皮气孔保卫细胞变大,叶绿体数变多,叶片变小,花径变大,舌状花数目减少,管状花数目增多。     

秋水仙碱诱导栝楼四倍体

采用改良L.D.Cua法诱导短把栝楼获得四倍体,研究了0.2%浓度的秋水仙碱诱导短把栝楼染色体倍性变异的最佳处理时间、四倍体栝楼与原栝楼的形态特征及其四倍体与二倍体的细胞学鉴定。结果表明：0.2%秋水仙素的最佳处理时间为72 h,诱变率达82.1%。与二倍体相比,四倍体表现为：生长变缓;叶片增大、皱缩和变厚、叶形指数减小;叶片保卫细胞增大,气孔密度降低,叶绿体数目增加,叶色加深。根尖压片表明：二倍体的染色体数目为2n=66,大部分四倍体植株的染色体数目为2n=132,M2代部分四倍体植株也存在嵌合现象。     

秋水仙素诱导蝴蝶兰原球茎产生多倍体研究

为探究秋水仙素处理对蝴蝶兰染色体加倍的效果,本研究以蝴蝶兰杂交组合H-03(2n=2x=38)的原球茎为材料,采用共培法诱导多倍体,秋水仙素浓度分别为0.05%、0.1%、0.2%,处理时间分别为5、10、15 d。结果表明,0.05%浓度的秋水仙素处理15 d效果最佳,变异率为50.00%,变异株存活率为30.00%;对处理后的植株进行流式细胞仪鉴定和染色体压片分析,发现变异株存在大量嵌合体;与对照相比,变异株形态特征表现为植株矮化、叶色加深、叶片表面粗糙、叶片变大、叶片偏短或偏圆等,部分植株表现出植株畸形或叶片扭曲;气孔观测结果表明,变异株气孔偏圆,单位面积气孔数减少,气孔增大,长、宽较对照分别增长76.45%、38.99%。本研究结果为蝴蝶兰体细胞无性系变异新品种的培育提供了参考。     

大叶铁线莲四倍体的诱导及初步鉴定

以毛茛科铁线莲属植物大叶铁线莲为试验材料,采用初生分生组织处理法用0.2%秋水仙碱溶胶诱导大叶铁线莲顶芽以期获得多倍体。本研究探讨了秋水仙碱诱导大叶铁线莲的最佳处理时间,并对所得的诱变株进行形态学和细胞遗传学鉴定。结果表明:0.2%秋水仙碱诱导多倍体最佳时间为48h,诱导率可高达80%。形态学方面,所得变异株与对照株叶片相比,叶片加长、加宽、增厚,叶形指数变小。变异株的叶片下表皮气孔明显大于对照,而气孔密度明显小于对照;保卫细胞中的叶绿体数目明显高于其二倍体对照;SPAD值表明叶绿素含量也有明显增加。根尖有丝分裂细胞学鉴定结果表明:对照株的染色体数目均为16(2n=2x),而变异株的染色体数为32(2n=4x),由此表明本研究通过秋水仙碱诱导大叶铁线莲成功获得四倍体。     

黄毛草莓叶片离体再生及其同源四倍体的诱导

为探究黄毛草莓叶片离体再生率及其同源四倍体的诱导效果,以黄毛草莓试管苗叶片和离体芽尖为材料,研究了不同激素配比对叶片离体再生的影响以及不同秋水仙素浓度和处理时间对其染色体的加倍效果。结果表明,黄毛草莓离体叶片在MS+1.0 mg·L-1TDZ+0.1 mg·L-1IBA的培养基上不定芽的再生率最高,为73.19%;0.15%秋水仙素处理草莓离体芽尖48 h诱导效果最佳,诱导率达13.33%,获得了经染色体计数确定为四倍体的株系12个,四倍体基因组DNA含量发生了倍增,染色体数量由14(2n=2x=14)条增加到28(2n=4x=28)条。与二倍体相比,四倍体黄毛草莓叶片更大更厚,颜色更深,叶形指数减小,气孔变大,叶绿素含量升高,整体植株生长势较强。黄毛草莓同源四倍体株系的获得为黄毛草莓的进一步研究奠定了基础。     

秋水仙素诱导杂交兰四倍体及倍性鉴定

以杂交兰F1代无菌苗(2n=2x=40)为供试材料,分别用浓度为0.02%、0.05%、0.10%和0.20%的秋水仙素对其根状茎处理24、48和72h,成功获得了四倍体植株。试验结果表明,以0.10%的秋水仙素处理48h诱导效果最佳,变异率为36%。根尖染色体压片表明,四倍体染色体数为2n=4x=80,二倍体对照为2n=2x=40。加倍后的植株较二倍体植株粗壮,叶片变厚,颜色变深,茎基部粗壮,颜色深,生长势变缓,根系变粗,根状茎增粗,可作为新材料加以利用。     

苹果叶片离体培养中秋水仙素加倍效应的研究

用不同浓度的秋水仙素与２％二甲基亚砜的复合溶液处理苹果栽培品种嗄拉和千秋的试管苗离体叶片，结果表明，以０．５％秋水仙素溶液处理４天的效果最佳，诱变频率达５６．１％，诱变后获得的四倍体植株在形态学和细胞学上均发生了明显变异，与对照相比，变异株茎矮、粗壮、节间短、叶色浓厚，单位面积气孔数目减少，气孔体积增大，染色体数目加倍，２ｎ＝４ｘ＝６８，核仁数目增加，梢端组织学切片证实，获得的同质四倍体频率为８６     

二甲戊灵与秋水仙素离体诱导虎眼万年青多倍体发生的比较研究

以白花虎眼万年青(Ornithogalum thyrsoides)组培的丛生芽(2n=2x=12)为试验材料,采用溶液浸泡法,研究秋水仙素和二甲戊灵的不同浓度和处理时间对白花虎眼万年青多倍体的诱导效果,观察根尖细胞染色体数,叶片下表皮保卫细胞及叶片组织结构特征。结果表明:800μmol·L-1二甲戊灵浸泡24h,变异率达46.67%,0.05%秋水仙素处理48h,达42.22%,两种诱变剂诱导变异的差异虽不显著,但前者处理后材料存活率高于后者(前86.67%,后80.0%)。对变异的材料通过细胞学鉴定,发现两种诱变剂均能诱导出四倍体白花虎眼万年青。从处理时间短,材料死亡率低,对人畜伤害小,成本低,变异率较高以及两种诱变剂处理得到的多倍体气孔、叶片结构特征的相似性等综合条件考虑,可将二甲戊灵作为秋水仙素的替代品诱导多倍体。此外,对比二倍体与四倍体植株形态特征、叶片下表皮气孔数目及大小,可将气孔大小作为初步快速检测多倍体的有效方法。本试验用2种诱变剂诱导白花虎万年青多倍体,并从形态、气孔、叶片组织结构比较诱导前后差异,旨在为白花虎眼万青年多倍体新品种研究提供数据支持。     

高产、抗病同源四倍体不结球白菜黄心乌新材料的创制

不结球白菜黄心乌是长江中下游流域特别是安徽地区的主栽品种之一。为获得品质优良的黄心乌新材料,本研究使用0.2%(w/v)的秋水仙素溶液点滴处理二倍体黄心乌幼苗的子叶生长点,通过形态学和细胞学的方法筛选到同源四倍体黄心乌植株,并对二、四倍体黄心乌的农艺性状以及品质进行分析。形态学研究结果表明,四倍体植株的株型、叶片、花、种荚表现出巨大型;四倍体叶片的气孔变大,气孔密度变小;花粉粒体积比二倍体变大且呈现出棒状。细胞学分析结果发现,四倍体植株的根尖染色体数目是二倍体的2倍。流式细胞仪分析结果表明,四倍体植株的荧光强度为二倍体的2倍。营养品质方面,与二倍体相比,四倍体纤维素、有机酸含量显著上升,叶绿素、硝态氮含量显著下降,可溶性糖及可溶性蛋白含量无显著变化。此外,结合光响应曲线分析结果发现,强光条件下,四倍体对光强的适应能力更强。在抗病性检测中发现,四倍体的灰霉菌抗性明显优于二倍体。本研究获得了高产、抗病的同源四倍体不结球白菜黄心乌新材料,为不结球白菜育种工作提供了新的种质资源。     

矮紧型、抗病同源四倍体不结球白菜矮桩秋冠新材料创制

为创制矮紧型、抗病同源四倍体不结球白菜矮桩秘冠新材料,使用浓度为0.2%的秋水仙素处理二倍体不结球白菜矮桩秋冠植株子叶期生长点,通过形态学、解剖学、细胞学特征和使用流式细胞仪进行鉴定,筛选出同源四倍体植株,并进行农艺性状、营养指标、光合能力和抗病性测定。形态学鉴定结果显示,四倍体植株的种子、花器官、荚果表现出巨大化,但整体株型和叶片变小。解剖学鉴定结果发现,四倍体植株气孔变大但单位面积密度减小,花粉粒变大且形状不规则。细胞学鉴定中,二倍体植株细胞染色体数为20条,四倍体为40条,是二倍体的2倍。流式细胞仪鉴定结果表明,二倍体细胞核DNA相对含量峰值在50处,四倍体细胞核DNA相对含量峰值在100处,与细胞学鉴定结果一致。农艺性状中,四倍体植株的单叶质量、十叶厚有所增加,但叶柄长、株高、叶长等有一定程度的降低。营养品质中,四倍体可溶性糖、纤维素、叶绿素含量减少,氨基酸、有机酸含量增加。光合特性分析结果发现,与二倍体相比,四倍体的最大净光合速率、表观量子效率、暗呼吸速率分别增加了29.192%、16.071%和14.0947,光饱和点降低,说明四倍体植株比二倍体具有更强的光合效率。抗病测定结果发现,四倍体植株叶片对灰霉病的抗性明显增强。经选育获得了矮紧型、抗病不结球白菜同源四倍体矮桩秋冠新材料,为不结球白菜种质创新和新品种培育奠定了基础。     

木薯同源四倍体的离体诱导及鉴定

为获得木薯新品种四倍体种质资源,以木薯主栽品种南植199组培苗单芽茎段为试验材料,利用秋水仙素诱导同源四倍体,统计不同浓度秋水仙素对外植体成活率和诱导效果的影响,并对二倍体与同源四倍体进行形态学、解剖学和生理生化方面的比较。结果表明,外植体成活率随秋水仙素浓度升高而降低,0.05~0.10 g·L^-1是秋水仙素诱导木薯同源四倍体的适宜浓度范围。本研究获得了10个同源四倍体株系和1个嵌合体株系。与二倍体木薯相比,四倍体木薯叶片增厚,叶型变宽,保卫细胞增大,叶绿素含量增加。本研究拓宽了木薯种质资源,并为诱导鉴定木薯四倍体提供了一套高效的技术方法。     

Identification of double/twin bolled and bicolor unit ‘cluster’ in <Emphasis Type="Italic">Gossypium hirsutum</Emphasis> L. cotton

Cultivated tetraploid Gossypium hirsutum L. (Malvaceae) cotton has been characterized as having one flower at a primordia. In a strain (CSH-13) of Gossypium hirsutum cotton, four plants out of 103 had two to three double bolls or twin bolls/plant during 2003–2004 crop season at CICR regional station, Sirsa, Haryana, India. Progeny evaluation in 2004 and 2005 crop seasons indicated that plants raised from seeds harvested from double bolls/twin bolls produced plants bearing double bolls only and plant progeny raised from the seed of single bolls from these mutant plants produced two to three double bolls per plant similar to the parent. Progeny testing revealed that double boll formation is the result of spontaneous mutation and environment does not influence its expression. The mutant is early in maturity by 10–15 days, naked seeded and possesses comparable agronomic characters with normal plants. Another spontaneous mutant of Gossypium hirsutum having more than three appendages originating from primordia.: four bolls one leaf, three bolls one leaf, two bolls two leaves, one boll three leaves were obtained from the population of CISV-13 strain in crop season 2003. These appendage groups have been described as bicolor unit by Clemens Bayer, [Zur Infloreszenzmorphologie der Malvales. Dissertations Botany 212 (1994)] and cluster mutant by Russell and Luther [J Cott Sci 6 (2002) 115]. One to three bolls in a cluster or bicolor unit were observed without formation of seeds and lint. About one-third of the total bolls on the mutant plant were of this type. In progeny testing during 2004 and 2005 crop seasons, this mutant produced plants exclusively with more than three appendages revealing that plants identified in 2003 were due to spontaneous mutation. This mutant was also early and naked seeded. Many of the other characters of the mutant plants were comparable with the wild type plants of the strain. Both the mutants were observed having economic impact due to their better yielding ability as compare to respective parents.     

Ploidic and Molecular Analysis of ‘Morado de Huetor’ Asparagus (Asparagus officinale L.) Population; A Spanish Tetraploid Landrace

A study has been conducted into the genetic variation analysis of the tetraploid asparagus landrace ‘Morado de Huetor’ and its relationship with current commercial cultivars using Random Amplified Polymorphic DNA (RAPD) molecular markers. The presence of different ploidic levels in the ‘Morado de Huetor’ landrace has been also studied using cytogenetic analysis. Ten decamer oligonucleotides were used to obtain RAPD markers and to characterise 52 individuals of ‘Morado de Huetor’, 7 of the tetraploid cultivar ‘Purple Passion’, and 55 of 5 diploid hybrid commercial cultivars. Jaccard similarity index was calculated and a cluster analysis using UPGMA (Unweighted Pair-Group Method using Arithmetic Average) was performed. Tetraploid asparagus, ‘Morado de Huetor’ and ‘Purple Passion’, were well differentiated from the diploid ones showing a higher genetic variability. This result suggests a different origin for tetraploid and diploid asparagus varieties. Within the diploid cultivars a variety distribution was obtained. A specific monomorphic band (OPB20₈₈₃) was found in tetraploid varieties. Within the diploid cultivars, two bands (OPB20₈₃₀, OPC15₇₀₅) can be used to distinguish between the hybrid cultivars used in this study. Tri-, penta-, hexa-, and octoploid plants were found in ‘Morado de Huetor’. The origin of these ploidic levels is discussed and their use in the development of new varieties is proposed. In conclusion, ‘Morado de Huetor’ is a genetic resource that could be used to increase the narrow genetic background reported in diploid asparagus cultivars.     

High proportion of diploid hybrids produced by interspecific diploid × tetraploid <Emphasis Type="Italic">Sorghum</Emphasis> hybridization

A perennial version of grain sorghum [S. bicolor (L.) Moench] would create opportunities for greatly reducing tillage and preventing soil degradation. Efforts to select for perenniality and grain production among progeny of hybrids between S. bicolor (2n = 20) and the weedy tetraploid perennial S. halepense (L.) Pers. (2n = 40) are complicated in that F₁ hybrids produced by diploid × tetraploid sorghum crosses are usually tetraploid. In 2013, a set of random pollinations between 19 diploid cytoplasmic male-sterile inbred lines and 43 tetraploid perennial plants produced 165 F₁ hybrid plants, more than 75% of which had highly atypical plant, panicle, and seed phenotypes. Phenotypic segregation in F₂ populations derived from atypical hybrids was also anomalous. Examination of mitotic metaphase cells in F₁ or F₂ root tips revealed that 129 of the 165 hybrids were diploid. Parentage of the diploid progenies was confirmed using simple-sequence repeat analysis. The mechanism by which diploid hybrids arise from diploid × tetraploid crosses is unknown, but it may involve either production of monohaploid (n = 10) pollen by the tetraploid parent or chromosome elimination during early cell divisions following formation of the triploid zygote. The ability to produce diploid germplasm segregating for S. bicolor and S. halepense alleles could have great utility, both for the development of perennial sorghum and for the improvement of conventional grain sorghum.     

Studies on awns in polyploid rice (Oryza sativa L.) and preliminary cross experiments of a special awnless tetraploid rice

Awns are an important genetic character of rice (Oryza sativa L.) and almost all awnless diploid varieties will change into awned varieties after polyploidization. In order to increase the understanding of awns and promote the development of polyploid rice breeding, the effect of polyploidization on awns was investigated and the development and distribution characteristics have been summarized. Furthermore, cross experiments were made between a special awnless tetraploid rice and two awned tetraploid rice. The results showed that the awns of polyploid rice initially emerged at the pistil–stamina formation stage and there was a stable corresponding relationship between awn development and other organs or development characteristics. Polyploidization had a great effect on awns. Awn length was considerably affected when diploid rice was doubled to polyploid. However, the degree of change varied between varieties. The F₁ hybrids between awnless and awned tetraploid rice in this research were all partial awn types and showed that the larger the mean, maximum awn length and the percentage of awned spikelets (PAS) of their awned parents, then the larger these of their F₁ hybrids. But different cross combinations showed some variation. The F₂ generations were still all partially awned and awnless type was not found. And the awn length and PAS of the plants developing from awned grains were significantly greater than that of the plants developing from awnless ones. This showed that the awn character of a grain could affect the plants developing from this grain. The results laid the foundation of further research of awn inheritance.