工业大麻雄性相关RAPD和SCAR标记的筛选与鉴定

利用42条RAPD（Random Amplified Polymorphic DNA）随机扩增引物分析工业大麻品种“火麻一号”组成的雄性或雌性DNA池（DNA pools）,结果显示,引物OPV-08扩增得到一条大小为869bp与工业大麻雄性相关的特异条带。根据测序结果,合成了两条SCAR（Sequence Characterized Amplified Region）标记引物,该SCAR标记不仅可以对工业大麻雌雄异株材料花期已知性别的雌雄植株进行准确鉴定,还可以对幼苗期未知性别的大麻雌雄植株进行鉴定;也可对雌雄同株材料可能出现的雄化进行早期鉴定。这不仅为工业大麻早期性别鉴定提供基础,且为减少雌雄同株材料的雄化提供支撑。     

苦瓜性别相关SSR和SRAP分子标记的筛选及分析

为揭示苦瓜雌雄花芽基因组间的分子差异,参照BSA法原理,构建了普通苦瓜雌雄花芽的DNA池和不同性型苦瓜雌雄花芽的DNA池,从26对SSR引物和30对SRAP引物中筛选特异引物对这些基因池进行差异性扩增。分析结果发现,两种标记分别筛选得到9对SRAP特异引物和8对SSR特异引物,引物有效率为30%和30.8%。苦瓜雌花芽和雄花芽DNA序列存在差异,SSR标记在分析普通苦瓜花芽DNA池和不同性型苦瓜花芽的DNA池时,多态性比率分别是10.7%和22.5%,SRAP的多态性比率则是7.4%和24.7%。SRAP扩增的雌性花芽的特异条带数较多,SSR分析雌雄花芽却只扩增出雄花芽的特异条带。但这些条带是否与性别表达基因紧密连锁,还需利用不同苦瓜的雌雄花芽DNA进行验证。这一研究结果为揭示苦瓜性别表达的分子机理奠定了基础。     

雌雄同株黄檗有性繁殖特性研究

一般认为黄檗为严格的雌雄异株植物,但本研究首次发现自然界存在两性植株。为了研究两性植株的有性繁殖能力,以两性植株为试材,雌株和雄株为对照,对两性植株性器官形态和繁殖能力进行了研究。通过2年对5000余朵花的研究发现：（1）两性植株的花粉和柱头与雄株和雌株在形态和发育上没有差异,花粉的萌发率在23%以上,与雄株花粉基本一致;（2）两性植株花粉授粉至雌株后得果率为86.69%,两性植株雌花在接受雄株花粉后的得果率为91.30%,自然受粉条件下的得果率为95.69%,自交授粉的得果率为88.01%。两性植株雄性和雌性都能正常繁殖,也可以成功自交繁殖。本研究为黄檗性别分化研究提供了新方向,为其生物保护提供了新途径。     

亚麻显性雄性核不育基因的RAPD标记

应用252条10-mer随机引物对遗传背景相似的可育株和不育株亚麻进行了RAPD分子标记,在不育株与可育株之间寻找DNA的多态性差异带,结果发现,在252条随机引物中有2条引物(即S62和S135)可分别得到1个与显性核不育的雄性基因有关的RAPD分子标记为S62-500和S135-350。     

栝楼雌雄株苗期、花期生理生化指标差异的比较研究

研究栝楼雌雄株不同发育时期植株相关生理指标的变化,为栝楼植株早期性别判定提供参考。通过定株标记,测定栝楼苗期和花期雌株雄株叶片中的可溶性糖、水溶性酚、内源激素以及核酸的含量。水溶性酚、核酸、生长素(IAA)和赤霉素(GA)的含量随着植株的生长而降低,可溶性糖、脱落酸(ABA)、玉米素(ZR)的含量则随着植株的生长而增加;苗期栝楼雌株叶片中可溶性糖、ABA和ZR的含量均显著高于雄株,水溶性酚和核酸的含量则显著低于雄株,GA的含量在雌雄株之间无显著差异。叶片中的可溶性糖、水溶性酚、核酸、ABA和ZR的含量可作为栝楼早期性别判定的参考指标。     

软枣猕猴桃雌雄株光合特性差异研究

为了比较软枣猕猴桃雌雄株间光合特性差异,以野生软枣猕猴桃成龄雌雄株为试材,利用CI-340便携式光合测定仪测定了光合特性。结果表明：软枣猕猴桃雌株的净光合速率日变化曲线为双峰型,而雄株为三峰型,雌雄株均出现“午睡”现象;软枣猕猴桃雌株蒸腾速率日变化曲线是单峰型,而雄株为双峰型,雌雄株“午睡”现象不太明显;雌株气孔导度日变化曲线为双峰型,而雄株为三峰型;雌雄株细胞间隙CO2浓度日变化曲线趋势基本一致,均出现3次高峰,但出现的时间和峰值具有显著差异;雌雄株叶面温度日变化均为双峰型,但高峰出现的时间和峰值均有较显著差异。软枣猕猴桃雌雄株间光和特性日变化在趋势和数值上均具有显著差异,可以作为性别鉴定的有效方法。     

DNA分子标记在银杏中的应用

DNA分子标记广泛用于生物研究的许多方面。本文简要介绍了RFLP、RAPD、AFLP、SSR及ISSR等几种目前常用分子标记的原理,归纳总结了分子标记在银杏中的应用研究进展。(1)获得了2个雄性特有的RAPD标记和2个雌性特有的AFLP标记,为银杏的早期性别鉴定及相关基因克隆奠定了基础;(2)采用RAPD标记和ISSR标记对我国部分栽培品种进行了分子鉴别和分类的研究,编制了一些品种的DNA指纹检索表;(3)利用RAPD和ISSR标记对一些群体、个体及栽培品种或变异类型进行了遗传分化和遗传多样性研究,结果发现银杏具有较高的遗传多样性,群体间、个体间、栽培品种及类型间都存在不同程度的遗传分化;(4)利用ISSR标记对一些个体的遗传杂合性进行了研究,结果显示个体的平均杂合率为43.53%;(5)构建了包含62个RAPD标记、19个连锁群的银杏分子遗传图谱;(6)探索了银杏优先保护种群的确定。分子标记在银杏其它方面的应用还很少。今后,除了继续对上述方面进行深入系统的研究外,还应充分运用DNA分子标记技术,开展银杏的分子标记辅助选择育种、种质评价与鉴别及保育生物学等方面的研究。     

为何金弹子开花不结果?

金弹子属柿树科,柿树属,其植物习性为雌雄异株,这是单株种植金弹子开花不结果的主要原因。不论你种植的是雄株还是雌株,在4月分开花时因缺少雄性花粉或雌性子房而无法开花结果。为使金弹子挂果,可以采用雌雄株同时种植的方法,能使雌株挂果。在开花期采取人工授粉的方式,更能大大的提高座果率。如要使雌雄植株都能开花结果,就可以采用     

乡村驿站

什么是DNA分子标记指纹图谱鉴定什么是DNA分子标记指纹图谱鉴定DNA分子标记指纹图谱鉴定是根据任何品种都具有该品种所特有的细胞遗传学的特征,利用分子标记技术,如限制性片段长度多态性(简称RFLP)、     

海岛棉抗黄萎病基因SSR标记研究

以高抗黄萎病的海岛棉品种Pima 90-53和高感黄萎病的陆地棉品种中棉所8号的182个F2单株为标记群体,在田间病圃中鉴定F2单株,并通过培养室人工接菌法鉴定F2:3家系,以进一步确定相应F2单株的抗病性,经χ2c适合性测验,抗病、感病植株比例符合3∶1。采用混合分组分析法(BSA)对768对SSR引物进行筛选,发现引物BNL2440和BNL3255在抗、感DNA池呈现多态性,其中BNL3255在抗、感DNA池之间扩增出一条大小为208bp的多态性片段,定名为BNL3255-208。以Mapmaker/Exp(Version3.0b)软件分析F2单株检测结果,BNL3255-208标记与棉花黄萎病抗性位点之间的遗传距离为13.7 cM。     

石刁柏性别表现与同工酶的关系

采用聚丙烯酰胺凝胶电泳对石刁柏（ＡｓｐａｒａｇｕｓｏｆｆｉｃｉｎａｌｉｓＬ．）雌雄植株不同器官及不同组织的过氧化物酶同工酶谱进行了研究，结果表明，除根部外，尽管鳞片、茎尖、拟叶等器官酶谱有明显差异，但雌雄株间差异有相同的趋势。雄株均比相应的雌株少一条酶带，雌雄植株组织培养获得的愈伤组织和茎尖过氧化物酶同工酶酶谱差异也有类似的规律。说明石刁柏性别差异与过氧化物酶同工酶的数目有关，过氧化物酶同工酶谱的差异可以作为性别鉴定的指标。     

雌雄石刁柏嫩茎的营养价值和品质的比较研究

本文对不同性别和不同部位的石刁柏嫩茎的营养价值和品质进行了比较研究。结果表明,雄石刁柏嫩茎在氨基酸、类胡萝卜素、铁和锌含量等方面均较雌的高,所以雄石刁柏嫩茎营养价值较高;雄石刁柏嫩茎的纤维含量较雌的低,所以雄石刁柏嫩茎品质也较好。然而,雌石刁柏嫩茎的可溶性糖、还原糖、脂肪和钙含量高于雄的;维生素Ｃ含量二者相近。同一性别的石刁柏嫩茎,营养价值和品质都是上部最佳,中部次之,基部最差。     

芦笋性别的化学鉴定研究

在成熟芦笋叶片与未知性别的芦笋叶片提取液中加入Ⅰ号裴林试剂和Ⅱ号裴林试剂,静置60分钟后,成熟芦笋雌株上清液呈翠绿色,雄株为黄绿色;而未知性别芦笋上清液颜色无区别,但沉淀量不同。     

Development of a rapid method for identifying asparagus super-males using, N-(4-chloro-2-trifluoromethylphenyl)-N'- propoxyacetamidine to induce flowering

In order to shorten the term for the identification of asparagus super-maleplants, a method usingN-(4-chloro-2-trifluoromethylphenyl)-N'-pro-poxyacetamidine (AM12) toinduce flowering was developed. This method is conducted as follows: seedsfrom andromonoecious flowers are treated with AM12 to induce flowersto form on the seedlings, the new male flowers are crossed with normalfemale flowers, and the progeny seeds are again treated with AM12 toidentify super-male plants from the sex ratio of the seedlings.Asparagus officinalis cv. `UC157' seeds were treated with AM12.The seedlings were induced to flower at a high frequency in 25 days. Thesex ratio of the plants was 1:1 and the male flowers had the pollengermination ability. One male flower induced by AM12 was crossed withfemale plant. This cross produced many progeny seeds. The seeds weretreated again with AM12, and induced to flower. Two super-male plantswere found among the progeny of andromonoecious flowers of the all-malecultivar `Gijnlim' within six months by this method. This method thusshortens the time for identifying super-males. Since the female flowers werefertile as well, AM12 treatment would also be effective for cross breeding.     

Sex distinction of asparagus by loop-mediated isothermal amplification and observation of seedling phenotypes

Asparagus (Asparagus officinalis L.) is a dioecious plant. In general, male and female plants are used for open-field culture and intensive cultivation, respectively. Farmers distinguish between the sexes by observing the form of the flower organs. However, because flowering begins 2?C3 years after planting, the sexes cannot be differentiated at transplantation by using this method, and planting of an all-male population is not possible. In this study, the usefulness of loop-mediated isothermal amplification (LAMP), a simple method of gene amplification, for sex distinction at the DNA level was determined. In addition, the phenotypic differences in seeds and seedlings of male and female plants were investigated for application as a method of early sex distinction. By using the LAMP method, the sex could be correctly identified in 100% of the seedlings, suggesting that this method is effective for sex distinction at the gene level. Principal component analysis was conducted with 11 selected parameters after investigating the seeds and seedlings of both male and female plants. The results revealed that male plants tend to have many stalks or cladophylls and female plants tend to have large plant forms, suggesting that the sexes can be distinguished by the external appearance of the seedlings before planting. LAMP and observation of the seedling phenotypes could be useful methods of sex distinction for increasing the efficiency of asparagus breeding.     

A method for sex identification in asparagus using DNA from seeds

Asparagus (Asparagus officinalis L.) is a dioecious species, with both male and female individuals. Male plants are more desirable to cultivate than female plants because they have higher yields, and, unlike female plants, they do not have a weed problem resulting from fallen seeds. A male-specific DNA marker is currently available to identify the sex of asparagus individuals using total DNA extracted from cladodes and roots. However, no published method is currently available for DNA extraction and PCR amplification from asparagus seeds. In this study, we tested several heat-resistant DNA polymerases for PCR and several methods for extracting DNA from asparagus seeds and successfully established a method for identifying the sex of asparagus seeds using this male-specific DNA marker. We found that PCR amplification of DNA extracted from asparagus seeds using simple methods such as single-step DNA extraction requires the use of high efficiency DNA polymerase. By contrast, many types of heat-resistant DNA polymerases can be used for PCR amplification of high-quality DNA extracted from asparagus seeds using a commercially available DNA extraction kit. Our method for sex identification of asparagus seeds could facilitate quality checking of all-male asparagus seeds and accelerate the screening of super-male asparagus.     

A new DNA marker for sex identification in purple asparagus

Garden asparagus (Asparagus officinalis L.) is an economically-important perennial crop. This plant is dioecious, as there are both male and female individuals; male individuals are preferred over females for agricultural production. To reduce the time required for garden asparagus breeding, various male-specific DNA markers are utilized. Male-specific DNA markers, such as Asp1-T7sp and MSSTS710, are currently available for sex identification in many asparagus cultivars. In the current study, we found that these markers are not suitable for sex identification in the purple asparagus cultivar ‘Pacific Purple’, as male-specific amplification of this marker was detected in some male individuals of this cultivar but not in other males. The Asp1-T7sp marker is suitable for use in sex identification in various Asparagus species related to A. officinalis, indicating that the region around this marker is conserved among these species. Thus, we isolated a DNA fragment around this marker by inverse PCR and produced a new DNA marker, MspHd, based on this sequence. However, like Asp1-T7sp and MSSTS710, MspHd was not suitable for sex identification in the cultivar ‘Pacific Purple’. Since all ‘Pacific Purple’ males have morphologically similar male flowers with functional stamens, we produced a new male-specific marker based on the sex determination gene, MSE1/AspMYB35/AspTDF1, which is responsible for stamen development. This marker, named AspMSD, is suitable for sex identification in ‘Pacific Purple’. In addition, this marker can be utilized for sex identification in various asparagus cultivars and some related Asparagus species.     

影响芦笋产量性状的相关性分析

对芦笋的6个农艺学性状指标进行相关性分析,并对产量和一级笋率进行了多重比较。结果表明,影响芦笋产量的重要因子有茎粗、株高和茎数。影响芦笋一级笋率的重要因子有茎粗、株高、产量。植株越高,产量越高,一级笋率也越高。茎粗、株高、茎数构成芦笋产量、品质特性,同时也是新品种选育的必备性状。通过对产量和一级笋率的多重比较发现,全雄品种在产量、一级笋率和其它农艺性状方面明显优于其它常规种,因此芦笋全雄品种的选育已成为目前芦笋育种的重中之重。     

RAPD and SCAR markers linked to the sex expression locus M in asparagus

Bulk segregant analysis (BSA), random amplified polymorphic DNA (RAPD) and sequence characterized amplified region (SCAR) methods were used to map molecular markers to the sex locus M of asparagus. Two parents, A19 (male, Mm) and MW25 (female, mm), and 63 progeny were used for the study. Two DNA bulks, one male and one female, were made by pooling equal amounts of DNA from 10 randomly selected progeny of each sex type. A total of 760 arbitrary decamer oligonucleotide primers were used for RAPD analysis. Primer OPC15 produced two RAPD markers, OPC15-98 and OPC15-30, both of which were linked to the M locus at a distance of 1.6 cM. Subsequently, amplified RAPD fragment OPC15-98 was cloned and sequenced. The sequence was then used to design flanking 24-mer oligonucleotide SCAR primers SCC15-1 and SCC15-2. Both of these SCAR primers amplified a single 980 bp fragment; the same size as the cloned RAPD fragment. However, the SCAR marker was dominant as was the original OPC15-98 band from which it was derived. These RAPD and SCAR markers could be used for scoring male and female progeny in the mapping population, but were not found to be applicable to other asparagus germplasm studied. This revised version was published online in July 2006 with corrections to the Cover Date.