Mitosis Observation on the Tetraploid Lilium oriental and 2n Gamete Hybrid Offspring of Lilium oriental

对东方百合四倍体正常植株,变异植株,2n配子杂种后代进行有丝分裂观察,结果表明四倍体正常植株的有丝分裂均正常而变异植株出现染色体桥现象;2n配子杂种后代多数植株有丝分裂正常,少数植株的有丝分裂极不正常出现了染色体桥和染色体落后,双核及多极分裂现象,从而产生染色体数目的多样性。     

Isozyme electrophoretic characterization of 29 related cultivars of lily (Lilium spp.)

Isozyme banding patterns (IBPs) were studied for cultivars of lily (Lilium spp.) by means of horizontal starch‐gel electrophoresis (SGE). An array of continuous histidine‐citrate buffer systems at eight ranges of pH and four extraction buffers were tested. On the basis of this survey, the extraction buffer two (Eb‐2) and the buffer system E at pH 7.7 were found to be suitable for detection of lily isozymes. Using the SGE technique, IBP in catalase (CAT; EC 1.11.1.6), esterase (EST; EC 3.1.1.1), malate dehydrogenase (MDH; EC 1.1.1.37), malic enzyme (MAL; EC 1.1.1.40), peroxidase (POX; EC 1.11.1.7), phosphoglucomutase (PGM; EC 2.7.5.1), phosphoglucose isomerase (PGI; EC 5.3.1.9) and 6‐phosphogluconate dehydrogenase (PGD; EC 1.1.1.44) were assayed. In total 29 cultivars were tested in this study: nine were analysed for all eight enzyme systems, 16 cultivars for seven systems, three for six, and one for five enzyme systems. Some IBP were identified as section‐specific biochemical markers. Eight enzymes systems were analysed by constructing a dendogram using the unweighted pair group method, arithmetic average (UPGMA) cluster analysis. The analysis indicated that the lily cultivars could be separated from other Lilium species, except for two L. x formonlogi cultivars:‘Hakuba’ and ‘Hakuko’ which could not be distinguished from each other by the isozyme patterns assayed here. This study shows that isozymes can provide useful biochemical markers for lily cultivar identification and to estimate the phylogenetic relationships among those cultivars.     

Genomic constitutions and flower characteristics of selected Aranda orchid cultivars

Summary Ten Aranda cultivars commercially grown in Singapore were selected to study their genomic constitutions and flower characteristics. Cytological evidence and breeding records of these cultivars showed that they are of three genomic classes. Four of them are diploid with AV genomes (one Arachnis and one Vanda genome), another four are triploid with AVV genomes and the remaining two are tetraploid with AVVV genomes. Sizes of flowers as well as of sepals and petals generally show significant increases from diploid to tetraploid. This trend reflects the increasing influence of Vanda resulting from additional one and two Vanda genomes in triploid and tetraploid respectively as compared to diploid cultivars. Among the three genomic classes, diploid cultivars generally bear less flowers per spray than those of triploid and tetraploid although exceptions may occur. There is no clear trend in the length of inflorescences although diploid cultivars tend to have less compact spray with flowers more distantly spaced out.     

不同百合品种间杂交亲和性研究

以国外引进的性状优良的栽培品种7个作为亲本进行杂交,摸索不同品种间交配亲和性规律,提高育种效率。试验结果表明：4种杂种系不同品种百合系内杂交,后代平均有胚率最高的为OO杂种系（43.63%）,其次为AA杂种系（25.35%）。以OO杂种系的‘多顿’、‘蒙特祖玛’为母本,与其他杂种系杂交,后代平均有胚率高与其他组合,亲和性好。‘多顿’ב穿梭’这个组合的结实率为50%,有胚率为50.8%,杂交亲和性最好。     

淡黄花百合根尖染色体C-带分析

利用Giemsa C-带方法对淡黄花百合(Lilium sulphuFeum Baker)进行了研究.结果表明:淡黄花百合(Lilium sulphureum)的染色体数目为2n=2x=24,每条染色体上都显示出特征带,带纹的深浅差异明显,其带 .型公式为:2n=24:2CI+2I+4CI++2CI++8I++2I+T++2IT++2I+N.染色体F有两条强弱不同的中间带和一条次缢痕带.通过Cdemsa C-带方法可以将淡黄花百合(Lilium sulphureum Baker)的每条染色体区分开.     

一种新铁炮百合花瓣总RNA的提取方法

本研究以新铁炮百合花瓣为材料,采用非异硫氰酸胍提取法、一步快速热酚抽提法、Trizoi试剂法、天为时代植物RNA抽提试剂盒法、Trizol试剂法与天为时代的植物RNA抽捉试剂盒相结合的改良法,提取花瓣总RNA.结果表明:Trizol试剂法与天为时代的植物RNA提试剂盒相结合的改良法是一种经济、有效的新铁炮百合花瓣总RNA的提取方法,能有效地抑制酚类物质、多糖等次生代谢产物对RNA的影响,可从新铁炮百合花瓣中获得质量高、完整性好的总RNA.提取的RNA经紫外光谱分析表明:A_(206)/A_(280)比值为1.722,电泳检测到28S RNA、18S RNA清晰的条带.反转录后双链cDNA的分子量大小分布区间介于0.1～5.0 kb之间,符合植物基因cDNA的长度范围,可满足下一步的分子实验.     

百合染色体细胞学研究进展

百合属绝大多数种是2n=24的二倍体,其核型具有稳定性,一般为3B型,存在少数多倍化现象,但广泛存在B染色体,百合染色体核型的差异正是环境因素和结构变异共同作用的结果。通过百合属C-带带型中单套染色体条带数及特征染色体可以清晰地区分形态学相似的百合属植物,但采用尿素法进行G带带纹的鉴定有更高的分辨率。原位杂交技术已运用于百合属植物的区分和杂种后代的鉴定,而GISH较FISH更适用于杂种百合的鉴定。     

麝香百合的抗热生理指标初探

以田间抗热表现不同的麝香百合品种White Forest和新铁炮百合基因型O2-28、O1-13作为试验材料进行38℃高温处理，测定百合叶片的可溶性蛋白含量、过氧化物酶活性、过氧化氢酶活性、蒸腾强度4个生理指标的变化，结果显示百合的可溶性蛋白含量均下降,抗热性越强的基因型下降幅度越缓慢。酶活性与抗热性正相关，抗热基因型的两种酶活性提高的幅度大大高于热敏感基因型。蒸腾强度的变化没有明显规律，与百合抗热无相关性。     

东方百合L_sCCR1基因的克隆及表达分析

百合茎秆的挺度直接关系到百合花采收后的货架寿命等重要商品性状,而这些商品性状与植物茎秆中的木质素的含量高低有关.本文应用RLM-RACE技术,从东方百合索蚌植株中克隆了木质素合成相关基因一肉桂酰辅酶A还原酶,定名为LsCCR1.该基因cDNA全长为1 499 bp,包括完整的阅读框,编码388个氨基酸.多重比对分析发现百合LsCCR1基因与其他植物上已经发现的CCR基因同源性多介于55%～66%之间.利用半定量PCR检测LsCCR1基因在百合不同组织中的表达差异,结果显示LsCCR1基因主要在百合的茎秆中表达,根部次之,叶片、鳞茎及花蕾中表达量较少,其表达模式与拟南芥、桉树及大麦中CCR基因的表达模式类似.     

高温胁迫对抽苔期新铁炮百合的生理影响

测定了2种新铁炮百合基因型K1-4和M2-9抽苔期的叶片在36℃高温胁迫下的耐热性生理变化。结果显示,田间表现较耐热的K1-4的叶绿素和可溶性蛋白下降缓慢,脯氨酸积累较多,过氧化氢酶（CAT）活性保持较好。表明脯氨酸与百合叶片耐热性呈负相关,其它3项生理指标与耐热性呈正相关。2种百合均是上部幼嫩叶片的耐热性比下部成熟叶片的强。     

22种百合属植物在青岛地区引种的适应性研究

为丰富青岛地区能够利用的百合属植物的优良种类,以荷兰引进的22个百合属植物品种为试材,对其在青岛地区的引种适应性进行研究,系统观测不同品系百合品种的物候期、生长规律、花部特征和花色参数等。结果表明,22个百合品种均能适应青岛地区的环境条件,不同品系百合花期集中在5月,亚洲百合杂种系[Lilium asiatica hybrids（AH杂种系）]、麝香百合杂种系[L. longiflorum hybrids（LH杂种系）]和麝香百合和亚洲百合杂种系间的杂交种[L. longiflorum× asiatic hybrids（LA杂种系）]平均单花花期分别为9.25、12.43、14.43天。不同品系百合株高随时间的延长而呈逐渐增高的趋势,但不同品系间增高幅度不同。不同品系百合品种花径、花苞数目和花量不同,LH杂种系的花径和花苞数目小于AH杂种系和LA杂种系。花色参数中,AH杂种系的明度参数L*低于其他品系,LH杂种系中‘Regale’红绿参数a*值最低,为-2.08,LH杂种系中‘Dancing girl’黄蓝参数b*值最低,为-4.61。研究结果明确了百合属植物在青岛地区的引种适应性,为进一步的合理开发利用提供理论依据。     

黄秋葵基因组DNA提取及鉴定

黄秋葵细胞内含有丰富的果胶类多糖等次生物质,降低了从中提取总DNA的产量和质量。为了获得高质量的基因组DNA,采用改进的十六烷基三甲基溴化铵（CTAB）法进行提取,并对总DNA进行了纯度和浓度的鉴定。结果表明,改良的CTAB法可有效去除次生物质对DNA的干扰,降低DNA的粘稠度,样品DNA的质量和纯度较高,可用于随机扩增多态性DNA（RAPD）扩增等分子标记分析。     

广东省野百合天然居群的随机扩增多态性DNA(RAPD)分析

本研究选择了中国广东省7个有代表性的山区作为野百合(Lilium brownii F.E.Brown ex Miellez)天然居群的采样点,对采集到的199份样品进行了随机扩增多态性DNA(RAPD)分析,以建立野百合天然居群RAPD分析方法体系以便后续研究,并了解广东省野百合天然居群多态性情况和居群内外个体及野百合...     

Identification and cytological analysis of 2n-pollen producers in Lotus tennis Wald. et Kit.

Two meiotic mutants of L. tennis (2n = 2x = 12) producing unreduced pollen are described. When crossed to male sterile L. corniculatus (2n = 4x = 24) plants, all progeny plants were morphologically similar to L. corniculatus, had 2n = 24 chromosomes, and in the cross, were fully compatible with L. corniculatus, indicating that the male parent plants were 2n-pollen producers. One of them also had ‘giant’ pollen grains. In metaphase II of both genotypes, there were parallel and tripolar spindles leading to dyad and triad formation, the latter being found most frequently. Since both the above-mentioned mechanisms result in first-division restitution-type microspores, the genotypes examined could be useful in breeding Lotus.     

亚洲百合鳞茎休眠过程中需冷量的研究

以亚洲百合（白天使,普瑞头）为材料,研究了不同低温及不同时间对百合鳞茎解除休眠的效应和百合鳞茎解除休眠的最适需冷量,结果表明：亚洲百合打破休眠的最有效温度为5℃,15℃以上对打破休眠无效,仅白天使对-2℃低温有一定感应, 对于0℃、10℃低温,白天使经0℃处理打破休眠效率高于10℃,普瑞头则与之相反;两种百合鳞茎解除休眠的最适需冷量分别为：白天使,1200C.U,普瑞头,840C.U。     

商陆抗病毒蛋白基因在麝香百合中的转化和表达

以麝香百合叶片愈伤组织为受体,利用根瘤农杆菌介导法将美洲商陆蛋白（PAP）基因和抗卡那霉素筛选基因以共转化的方式转入百合叶片愈伤组织中,然后在含有MS培养基中筛选愈伤组织并得到再生植株,在建立的农杆菌转化百合的遗传转化体系中,获得49株再生植株,经PCR检测表明PAP基因已经转移到有抗性愈伤组织再生出百合植株中。     

Production of diploid and triploid interspecific hybrids between Lilium concolor and L. longiflorum by in vitro ovary slice culture

Successful hybridization between Lilium concolor and Lilium longiflorum has not been reported but ovary slice culture technique, after cut-style pollination has now been used to produce diploid and triploid interspecific hybrids between these species. Reciprocal crosses between diploid cultivars (2n= 2x= 24) were conducted. 5, 10, 15, 20, 25, 30, 35, 40, and 45 days after pollination (DAP), ovaries were sliced and cultured on a modified Murashige and Skoog (MS) medium without growth regulators and NH₄NO₃, supplemented with 6% sucrose, 50 mg/1 yeast extract and 0.25% gelrite at pH 6.3. For the L. concolor × L. longiflorum cross, embryo germination was found to be best at 20 DAP, while for the L. longiflorum × L. concolor at 25 DAP. After transfer to a MS (half-strength) medium supplemented with 1.5% sucrose, 0.25% gelrite and 0.2% active charcoal at pH 5.8, diploid and triploid hybrid plants were developed. All regenerated plants were identified as hybrids on the basis of karyotype and isozyme analyses. Ovary slice culture technique as a method of producing polyploids is discussed.     

Genome classification of banana cultivars from South India using IRAP markers

Summary The banana cultivars are originated from the intra- and inter-specific hybridization of two wild diploid species, Musa acuminata Colla and Musa balbisiana Colla, contributing the A and B genomes, respectively. They are classified into genomic groups by scoring morphological features. Molecular markers provide a quick and reliable system of genome characterization and manipulation in breeding lines. In the present study a PCR based molecular marker specific for B genomes is been reported. The IRAP primer, designed based on the LTR sequence of banana Ty3-gypsy-like retroelement (Musa acuminata Monkey retrotransposon, AF 143332), was used to identify the B genome in the banana cultivars. Further a primer pair designed from B specific bands of Musa balbisiana `Pisang Gala' was used to classify AAB and ABB cultivars in the collection. Among the 36 cultivars tested with this primer, the B specific band was absent in the AA and AAA cultivars (except in one AAA and AAB cultivar) but present in all other AB, AAB and ABB cultivars. Among the triploid AAB/ABB, the PCR products with B specific primers showed restriction pattern polymorphism with AluI. In ABB genomes the band intensity was high whereas low intensity band observed in AAB genomes. Four cultivars reported to have the ABB genome showed a pattern similar to AAB, and one cultivar reported to have AAA genome showed a pattern similar to ABB genome, suggesting missampling or misidentification. The primers used in this study are useful to identify the presence of B genome in banana cultivars, and band intensity may be a preliminary indicator of ploidy level of the B genome but needs further studies with competitive PCR for clarification. These authors contributed equally in this paper.     

A tetraploid hybrid plant from 4x × 2x crosses in Vitis and its origin

To study the origin of unreduced (2n) gametes in diploid Vitis cultivars, we surveyed the occurrence of tetraploid hybrid seedlings from 40 interploid crosses with five tetraploid and seven diploid cultivars. A total of 250 seedlings from the interploid crosses were established through embryo culture and by seed sowing. In 20 2x × 4x crosses, no tetraploid hybrid seedlings were derived from 8,902pollinations. In 20 4x × 2x crosses, two tetraploid hybrid seedlings were obtained from 8,057 pollinations. Investigation of isozyme genotypes of the two tetraploid seedlings using three variable enzyme systems indicated that one of the two seedlings resulted from the union of a diploid egg with a 2n male gamete and that failure of second meiotic division resulted in the formation of the 2n male gamete. The percentage of giant pollen grains in `Muscat Bailey A', a pollen parent of the tetraploid seedling, was relatively high(about 5.9%) and 10.9% of the giant pollen grains germinated on agar medium. These results suggested that 2n pollen of diploid cultivars are useful for breeding tetraploid grape. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genomic constitution of Festulolium cultivars released in the Czech Republic

Genomic in situ hybridization (GISH) was used to characterize the chromosome constitutions of individual plants from a set of tetraploid and hexaploid cultivars of Festulolium developed and released in the Czech Republic from hybrids of Lolium multiflorum with Festuca pratensis and F. arundinacea. A simplified GISH protocol readily discriminated parental genomes in the hybrids and facilitated the screening of large numbers of plants per accession. The contribution of parental genomes in the cultivars tested ranged from predominance of chromatin from one of the parents to a more balanced contribution from both parents. However, in none of the cultivars were equal proportions of chromatin from both parents present. The parental contribution to the hybrids was both in the form of complete chromosomes or as chromosome translocations. In hexaploid cultivars from (L. multiflorum × F. arundinacea) × F. arundinacea hybrids the average numbers of complete L. multiflorum chromosomes ranged from 4.95 to 7.5 and the numbers of translocations from 6.33 to 10.21. Two tetraploid cultivars from (L. multiflorum × F. arundinacea) × L. multiflorum hybrids showed a strong prevalence of L. multiflorum chromatin and intergeneric translocations were rare. In the tetraploid cultivar ‘Perun’ of the L. multiflorum × F. pratensis hybrid there were 11.7 chromosomes of L. multiflorum and 14.7 recombined chromosomes on average. Reasons for the domination of one of the parental genomes in hybrid cultivars are not clear and are only partially explained by breeding history. Recombination rates of individual genomes in hybrids involving F. arundinacea were evaluated in double hybridization experiments. The results indicated a strong affinity of the L. multiflorum genome for the F. pratensis genome present in F. arundinacea and little affinity for the F. glaucescens genome. This suggests that introgressions from F. arundinacea into L. multiflorum are primarily limited to the F. pratensis genome which can be more readily accessed in L. multiflorum × F. pratensis hybrids.