Analysis of genetic diversity and structure across a wide range of germplasm reveals genetic relationships among seventeen species of Malus Mill. native to China

China is a center of diversity for Malus Mill. with 27 native species including 21 wild species and six domesticated species. We applied a set of 19 simple sequence repeat markers to genotype 798 accessions of 17 species (12 wild species and five cultivated species) of Malus originating from 14 provinces in China. A total of 500 alleles were detected. Diversity statistics indicated a high level of genetic variation as quantified by the average values of the effective allele number (N_e), expected heterozygosity (H_e), and Shannon’s Information Index (I) (10.309, 0.886, and 2.545, respectively). Malus sieversii (MSR; H_e=0.814, I=2.041, N_e=6.054), M. baccata (MBB; H_e=0.848, I=2.350, N_e=8.652), M. toringoides (MTH; H_e=0.663, I=1.355, N_e=3.332), and M. hupehensis (MHR; H_e=0.539, I=0.912, N_e=0.579) showed a higher level of genetic diversity in this study than the previous studies. MSR and MBB contributed to the origin and evolution of some accessions of M. domestica subsp. chinensis (MDC). However, other accessions of MDC showed a closer genetic distance with MBB and cultivated species, especially M. robusta (MRB), M. asiatica (MAN), and M. prunifolia (MPB). Not all accessions of MDC were descended from MSR in Xinjiang Uygur Autonomous Region of China. This research provides novel insights into the genetic relationships of Malus native to China, which will be useful for genetic association studies, germplasm conservation, and breeding programs.     

南方根结线虫诱发新疆野生樱桃李形成的根结形态和发育

以新疆野生樱桃李(Prunus sogdiana Vassilcz.)扦插苗为试材,通过人工接种南方根结线虫(Meloidogyne incognita)鉴定其抗性,同时对抗病与感病单株在接种后的酶活性变化以及根结的形成发育过程进行了研究.结果表明:新疆野生樱桃李对根结线虫抗性发生分离,大部分单株表现为感病,只有28％单株表现为抗病,并且其抗性与本试验室之前克隆得到的抗根结线虫相关基因psoRPM1的表达存在着相关关系.接种根结线虫后CAT、POD酶在抗病植株中的活性高于感病植株,说明它们可能参与了抗根结线虫的生理过程,而PPO和PAL活性则在接种后7～14日抗病植株高于感病植株,之后在两者之间差异不大.接种根结线虫后线虫由皮层进入中柱,并在10 d时观察到形成了大量多核的巨型细胞,随后巨型细胞经过发育到接种后22 d可观察到根系开始出现明显的根结.接种后30 d观察到巨型细胞出现空洞.     

豌豆属（Pisum）SSR标记遗传多样性结构鉴别与分析

 【目的】评价豌豆属（Pisum L.）2个种5个亚种下,共8个资源类群的遗传多样性水平,揭示豌豆属下资源群体结构及其遗传关系远近,验证传统植物学分类的可靠程度,为充分发掘、利用豌豆野生种质提供必要信息。【方法】利用21对豌豆多态性SSR引物,对来自世界5大洲62个国家的豌豆属94份栽培种质（P. sativum ssp. sativum var. sativum）及其1个近缘野生种（P. fulvum）,3个野生亚种（P. sativum ssp. abyssinicum、P. sativum ssp. asiaticum、P. sativum ssp.transcaucasicum）和3个野生变种（P. sativum ssp. elatius var. elatius、P. sativum ssp. elatius var.pumilio、P. sativum ssp.sativum var.arvense）的103份野生种质进行SSR标记遗传多样性分析;利用NTSYSpc2.2d软件估算其遗传距离,进行主成分分析（PCA）并绘制三维空间聚类图;利用Popgene V1.32估算种质群间的Nei78遗传距离等参数并进行UPGMA聚类分析,采用MEGA3.1绘制种质群间聚类图;采用Popgene V1.32估算种质群的等位位点分布等参数,利用Fstat V2.9.3.2进行种质群间遗传多样性差异显著性测验。【结果】21对豌豆多态性SSR引物共扩增出104条多态性带,每对引物平均扩增出4.95个等位变异,其中有效等位变异占65.56%;PSAD270,PSAC58,PSAA18,PSAC75,PSAA175和PSAB72等SSR引物最为有效。SSR等位变异在豌豆属植物学分类单位中分布均匀,但分类单位种质群间的遗传多样性在多数情况下差异显著。豌豆属野生种P. fulvum的遗传多样性远低于栽培种P. sativum;豌豆栽培种下,P. sativum ssp. sativum var. sativum和P. sativum ssp. asiaticum的遗传多样性最高,P. sativum ssp. elatius var. elatius和P. sativum ssp. transcaucasicum次之,P. sativum ssp. elatius var. pumilio、P. sativum ssp. sativum var. arvense和P. sativum ssp. abyssinicum最低。PCA分析发现,豌豆属种质资源由4个差异明显的基因库构成。“fulvum”基因库主要由野生种Pisum fulvum资源构成,“abyssinicum”基因库主要由栽培种下的P. sativum ssp. abyssinicum亚种资源构成,“arvense”基因库主要由栽培种下的P. sativum ssp. sativum var. arvense变种资源构成;“sativum”基因库由P. sativum ssp. asiaticum、P. sativum ssp. elatius var. elatius、P. sativum ssp. transcaucasicum、P. sativum ssp. elatius var. pumilio和P. sativum ssp. sativum var. sativum资源构成。“sativum”基因库构成豌豆栽培资源初级基因库;“fulvum”、“abyssinicum” 和“arvense”基因库共同构成豌豆栽培资源次级基因库。植物学分类单位间的Nei78遗传距离介于7.531～35.956,UPGMA聚类方法将豌豆属植物学分类单位聚成3个组群,“组群I”对应“sativum”和“arvense”基因库之和,“组群II”对应“abyssinicum”基因库,“组群III”对应“fulvum”基因库,聚类结果支持4个基因库的划分。【结论】豌豆属下多数植物学分类单位间遗传多样性差异显著,并分化成4个基因库。研究结果部分支持豌豆属下传统的植物学分类体系,并指出了其合理与不足之处。为拓宽豌豆育成品种的遗传基础,应充分发掘豌豆属下各基因库的遗传潜力。     

南方根结线虫与秀丽线虫同源基因的鉴定及其应用分析

南方根结线虫(Meloidogyne incognita)是一种分布广危害严重的植物寄生线虫,有关该线虫发育和侵染植物的分子机制报道较少;秀丽线虫(Caenorhabditis elegans)是第一个模式动物,对其功能基因组进行了详细而深入的研究,因此,利用生物信息学手段寻找和鉴定二者高度同源的基因可为深入阐明寄生线虫侵染植物的分子机制和防治提供重要的科学依据.本研究利用最新公布的南方根结线虫的9 165个Contig序列分别和秀丽线虫的361 207个EST序列、23 973个蛋白质序列进行比对分析,获得了2种线虫高度同源的基因,通过gene ontology分析2 793个高度同源基因表明,这些基因在调控线虫的生长和发育等过程中发挥重要作用,该项研究将为利用秀丽线虫基因功能资源进行寄生线虫基因功能研究和防治植物线虫病害提供重要的技术平台和新的策略.     

月季对根结线虫病抗性遗传研究

针对月季栽培过程中出现的最严重的根结线虫开门见山防治问题，开展了月季对根结线虫抗性遗传研究，研究结果如下：（1）对常用的4种月季砧木R.multiflora K1,K2,CE63及R.indica对左面种常见根结线虫即南方根结线虫（Meloidogyne incognita)、瓜哇根结线虫（M.javanica)、花生根结线虫（M.arenaria)及北方根结线虫（M.hapla)的抗性进行了测定，得出只有北方根结线虫能侵染月季，其余3种不能侵染。（2）根结指数与对数转化后的每克根中雌虫数、雄虫数、3-4龄幼虫数之和、2龄幼虫数、卵数、成虫有为3-4继幼虫数之总和、2龄幼虫与卵数之总和均呈线性相关关系，因此根结指数是进行抗性评价的可靠、简单及实用的指标，抗性划分标准如下：根结指数0.0-1.0，抗病；1.0-2.0，中等抗病；2.0-3.0，感病；3.0-4.0，比较感病；4.0-5.0，高度感病。（3）以根结指数为指标，通过对杂交所得74个子代的抗性评价试验表明；在2抗性亲本所得12个子代中，各子代抗性水平存在抗性分化，说明抗病基因不晨由单一显性基因控制；在所有的62个抗性亲本与感病亲本的杂交子代中，59个子代均比抗病亲本感病，表明抗病基因由隐性基因控制，因此，提出了月季对根结线虫的抗性遗传可能由2个基因决定。     

野生樱桃李对南方根结线虫的抗性

为探明野生樱桃李对南方根结线虫的抗性,以实生钵苗为试材,采用人工接种等方法,研究了野生樱桃李对南方根结线虫的抗性。结果表明：接种后30d,根中雌成虫数量占接种量的0.5%,依据抗性评价标准判定其高抗南方根结线虫;野生樱桃李对南方根结线虫的抗性存在显著的株间分离现象,表现为免疫、高抗、中抗和低抗4种类型,分别占群体总量的30.0%、52.5%、13.8%和3.8%;根内线虫数量为接种量的1.7%,雌成虫数量占根内线虫总数的29.4%,表明野生樱桃李对南方根结线虫具有极强的抗侵入作用和很强的抗发育作用,抗侵入是野生樱桃李对南方根结线虫的主要抗性机制。野生樱桃李是优异的抗南方根结线虫种质资源。     

AFLP初析小豆栽培和野生变种(Vigna angularis var.angularis and var.nipponensis)间演化与地理分布关系

 利用 12对AFLP引物 ,以饭豆标准品系M 0 0作对照 ,对来自中国、日本、韩国、尼泊尔、印度、不丹的 14 6份小豆栽培种 (Vignaangularisvar.angularis)和野生种 (Vignaangularisvar.nipponensis)种质的基因组DNA进行扩增 ,得到 313条多态性带。据AFLP多态性数据绘制的聚类图 ,可区分其中的 14 3份种质 ,表明小豆物种 (Vi gnaangularis)存在足够的遗传多样性 ,可用于资源材料的准确鉴别与分类。鉴于此 ,采用新开发的利用AFLP数据揭示核苷多样性的Innan’s进化树分析方法 ,进一步将 14 6份小豆资源分成 7个明显不同的地理演化群 ,即中国栽培种、日本栽培种、日本综合群 韩国栽培种、中国台湾野生种、中国野生种、尼泊尔 不丹栽培种和喜马拉雅野生种演化群。就上述地理演化群的遗传多样性、地理分布以及野生种与栽培种之间可能的演化关系进行了分析 ,初步认为栽培小豆至少应当有 4个不同类型的野生祖先和 3个不同的地理起源。     

Evaluation of Chinese rice varieties resistant to the root-knot nematode Meloidogyne graminicola

The root-knot nematode Meloidogyne graminicola, which is distributed worldwide, is considered a major constraint on rice production in Asia. The present study used the root gall index and number of nematodes inside the roots to evaluate resistance/susceptibility to M. graminicola in different subpopulations of Oryza sativa(aus, hybrid aus, indica, hybrid indica, temperate japonica, tropical japonica). Nematode development in highly resistant varieties was also evaluated. Analyses of randomly selected 35 varieties showed the number of M. graminicola nematodes inside the roots correlated very strongly(r=0.87, P≤0.05) with the nematode gall index, and the results from pot and field experiments revealed similar rankings of the varieties for resistance/susceptibility. Among the 136 tested varieties, temperate japonica displayed the highest gall index, followed by tropical japonica, indica, hybrid indica, aus, and hybrid aus. Zhonghua 11(aus), Shenliangyou 1(hybrid aus) and Cliangyou 4418(hybrid indica) were highly resistant to M. graminicola under both pot and field conditions. Further examination of nematode development suggested that compared to susceptible rice, M. graminicola penetrated less often into highly resistant varieties and more frequently failed to develop into females. The promising varieties found in the present research might be useful for the breeding of hybrid rice in China and for the further development of practical nematode management measures.     

榆中县黄瓜根结线虫鉴定及根系定殖效果测定

为了明确甘肃榆中县温室大棚黄瓜根结线虫种类及根际定殖情况,采用形态和分子生物学鉴定其种类,并室内测定和显微观察其在根际定殖效果。结果表明,甘肃榆中县黄瓜根结线虫不同虫态形态、大小和雌虫会阴花纹特征等均具有典型南方根结线虫(Meloidogyne incognita)形态特征。基于线粒体COI基因序列比对和构建系统发育树,鉴定发现榆中县黄瓜根结线虫线粒体COI基因序列与已报道的南方根结线虫台湾株系(KU517167)聚为一类,其相似性为99.75%;根结线虫多序列特异位点鉴定发现其与南方根结线虫孟加拉国株系(MN196556)和南方根结线虫印度株系(MN728697)聚为一类,相似性分别为100%和96.91%。经致病性测定发现,待接种根结线虫2龄幼虫30 d后其能够侵染黄瓜根系并形成大量根结,经染色发现黄瓜幼苗根系组织内寄生大量线形蠕虫、雌虫和卵囊,且镜检观察发现其形态与田间采集后分离的线虫形态一致。综上可知,引起甘肃省榆中县黄瓜根结线虫病的病原为南方根结线虫。     

桑树品种和种质资源干旱胁迫生长差异

通过对云南省目前生产上主栽的桑树品种和部分野生桑种质资源进行抗旱性试验研究.结果证明:栽培品种以云丰1号、云丰2号、丰田2号、丰田5号、强桑1号是目前抗旱性最好的品种组合,适合在云南推广,育种素材以山桑、蒙桑和鬼桑抗旱性最强,是最好的抗旱育种素材和云南裸山喀斯特地貌优势生态树种.     

Establishment and application of an SNP molecular identification system for grape cultivars

We aimed to develop a set of single nucleotide polymorphism (SNP) markers that can be used to distinguish the main cultivated grape (Vitis L.) cultivars in China and provide technical support for domestic grape cultivar protection, cultivar registration, and market rights protection. A total of 517 high-quality loci were screened from 4 241 729 SNPs obtained by sequencing 304 grape accessions using specific locus amplified fragment sequencing, of which 442 were successfully designed as Kompetitive Allele Specific PCR (KASP) markers. A set of 27 markers that completely distinguishes 304 sequenced grape accessions was determined by using the program, and 26 effective markers were screened based on 23 representative grape cultivars. Finally, a total of 46 out of 48 KASP markers, including 22 markers selected by the research group in the early stage, were re-screened based on 348 grape accessions. Population structure, principal component, and cluster analyses all showed that the 348 grape accessions were best divided into two populations. In addition, cluster analysis subdivided them into six subpopulations. According to genetic distance, V. labrusca, V. davidii, V. heyneana, and V. amurensis were far from V. vinifera, while V. vinifera×V. labrusca and V. amurensis×V. vinifera were somewhere in between these two groups. Furthermore, a core set of 25 KASP markers could distinguish 95.69% of the 348 grape accessions, and the other 21 markers were used as extended markers. Therefore, SNP molecular markers based on KASP typing technology provide a new way for mapping DNA fingerprints in grape cultivars. With high efficiency and accuracy and low cost, this technology is more competitive than other current identification methods. It also has excellent application prospects in the grape distinctness, uniformity, and stability (DUS) test, as well as in promoting market rights protection in the near future.     

油茶等植物提取物及制剂抑线效果

为了寻找植源性杀线虫活性材料,利用Bursaphelenchus xyluplilus 和Meloidogyne incognita 作为室内离体生测目标,测定了油茶籽饼等植物材料的杀线活性,以及茶皂素制剂和油茶籽饼对作物根结线虫的盆栽和大田防效。结果表明,油茶籽饼、茶果皮和茶籽粗提物均具有较强的杀线活性,当粗提物浓度为10 mg·mL^-1,作用72 h,B. xyluplilus校正死亡率分别为90%、100%和100%;作用48 h,油茶籽饼和茶籽粗提物对M. incognita达到中强杀线活性,其毒力有明显浓度效应;茶皂素制剂对番茄根结线虫病有良好盆栽防效,用茶皂素 （有效含量） 1.5和0.75 g·kg^-1处理盆土30 d后,番茄的根结和M. incognita 2龄幼虫（Juvenile 2, J2）均下降,对番茄苗期生物量有明显的增加。在瓜蒌出苗前1周,沟施油茶籽饼,用量2.5 t·hm^-2时对大田瓜蒌根结线虫病有较好的防效,显著抑制土壤中J2的增殖,对瓜蒌的挂果数和果径有一定的增加。该文首次报道油茶副产物对作物线虫病害的盆栽和田间防效,为直接利用油茶籽饼或茶籽等防治作物的根结线虫病害提供了依据。     

野生樱桃李对北方根结线虫和花生根结线虫的抗性

为探明野生樱桃李抗根结线虫种质资源的价值,以实生钵苗为试材,采用人工接种等方法,研究其对北方根结线虫和花生根结线虫的抗性.结果表明:接种后30 d,野生樱桃李根系中北方根结线虫、花生根结线虫的雌成虫数量分别占2龄线虫接种量的0.16％和0.03％,依据抗性评价标准判定其高抗北方根结线虫和花生根结线虫;接种北方根结线虫的群体中包含免疫、高抗和中抗3种类型,分别占群体总数的46％、48％和6％;接种花生根结线虫的包含免疫和高抗2种类型,分别占群体总数的56％和44％;接种北方根结线虫和花生根结线虫的植株未被侵染率分别为46％和52％,所有供试植株根系表面均未发现线虫卵块.野生樱桃李高抗北方根结线虫和花生根结线虫,其对北方根结线虫、花生根结线虫的抗性均存在显著的株间分离现象;抗侵入、抗发育和抗繁殖是其对北方根结线虫和花生根结线虫的主要抗性机制;野生樱桃李是抗根结线虫核果类果树砧木种质资源树种.     

Donors of resistance to potato golden nematode pathotype Ro1 derived from Solanum alandiae card

Clones of interspecific potato hybrids resistant to golden nematode have been created on the basis of wild species Solanum alandiae previously unused in breeding. The genetic nature of resistance is investigated by hybridological analysis and PCR marker technique.     

人工合成小麦对几种主要病害的抗性鉴定与评价

为了解人工合成六倍体小麦抗病性及其潜在利用价值,以95份人工合成小麦为试验材料,在人工接种全蚀病和条锈病、自然诱发白粉病和根腐叶枯病条件下,系统调查人工合成小麦对4种病害的田间抗病情况。结果表明,人工合成小麦中有较高比例的抗病材料,对全蚀病高抗以上材料占21%;对条锈病高抗以上材料占30%;对白粉病高抗以上材料占21%;对根腐叶枯病高抗以上材料占23%。CI93、CI108、CI187对全蚀病表现高抗、对其他3种叶部病害表现免疫,CI94对全蚀病、条锈病和白粉病表现高抗,对根腐叶枯病表现免疫,CI158、CI160、CI166对4种病害都表现高抗。     

The identification of presence/absence variants associated with the apparent differences of growth period structures between cultivated and wild soybeans

The cultivated soybean(Glycine max(L.) Merr.) was distinguished from its wild progenitor Glycine soja Sieb. Zucc.in growth period structure,by a shorter vegetative phase(V),a prolonged reproductive phase(R) and hence a larger R/V ratio.However,the genetic basis of the domestication of soybean from wild materials is unclear.Here,a panel of 123 cultivated and 97 wild accessions were genotyped using a set of 24 presence/absence variants(PAVs) while at the same time the materials were phenotyped with respect to flowering and maturity times at two trial sites located at very different latitudes.The major result of this study showed that variation at PAVs is informative for assessing patterns of genetic diversity in Glycine spp.The genotyping was largely consistent with the taxonomic status,although a few accessions were intermediate between the two major clades identified.Allelic diversity was much higher in the wild germplasm than in the cultivated materials.A significant domestication signal was detected at 11 of the PAVs at 0.01 level.In particular,this study has provided information for revealing the genetic basis of photoperiodism which was a prominent feature for the domestication of soybean.A significant marker-trait association with R/V ratio was detected at 14 of the PAVs,but stripping out population structure reduced this to three.These results will provide markers information for further finding of R/V related genes that can help to understand the domestication process and introgress novel genes in wild soybean to broaden the genetic base of modern soybean cultivars.     

安徽省3种中草药植物的根结线虫种类鉴定

白术、白芷和桔梗是3种重要的药用植物,它们的根茎有很高的药用价值。安徽省亳州市白术、白芷和桔梗这3种中草药植物种植田中发现有植株感染根结线虫病。为了明确感染这3种中草药的根结线虫种类,通过根结线虫雌成虫的会阴花纹、同工酶及分子生物学检测（rDNA-ITS区、28S rD2/D3区和rDNA-IGS区）进行了根结线虫种类鉴定。白术和白芷的根结线虫鉴定为花生根结线虫;桔梗的根结线虫鉴定为南方根结线虫。这是中国首次报道在白术和白芷上发现花生根结线虫病。这个结果为这3种中草药的根结线虫病防治提供了依据。     

Sequence Polymorphisms and Phylogenetic Relationships of hina Gene in Wild Barley from Tibet, China

The hina gene encodes a HINA protein in seeds of barley （Hordeum vulgare）, which was known to affect the grain hardness. 171 hina gene sequences from Tibetan wild barley accessions and worldwide were characterized. Across 1 452 nucleotides of 171 hina genes, 152 SNPs were detected, giving an average frequency of one SNP per 9.5 bases. There were 93 singleton variable sites （the nucleotide polymorphism only observed in a single accession）, 59 polymorphic sites （the polymorphisms found in two or more accessions） and 8 indels. A total of 18 haplotypes were defined, and most of the barley accessions shared one gene haplotype. H. spontaneum had a wider haplotype distribution. Through the analysis of median-joining network of the 18 haplotypes, 4 haplotype groups were found, which were testified by neighbor-joining tree based on the complete sequence alignment. Extremely low level of hina gene diversity was observed in Tibetan wild barley accessions, indicating that Tibet is unlikely a center of origin for cultivated barley.     

新疆野生樱桃李过敏性反应及其对南方根结线虫的抗性

以新疆野生樱桃李(Prunus sogdiana Vassilcz.)扦插苗为试材,接种南方根结线虫(Meloidogyne incognita)鉴定其抗性,并对抗病机理进行初步分析。结果显示:新疆野生樱桃李抗病植株发生过敏性(HR)反应,在接种后1d根系就出现典型的HR反应,并且在线虫周围细胞中发生HR反应的比例始终高于感病植株。在抗性植株发生HR反应的同时其体内大量产生过氧化氢,尤其在接种后1~2d过氧化氢和超氧阴离子的积累快速增加,这与HR反应特征基本一致。在接种南方根结线虫后,抗病植株中的VAD1和PO2表达均高于感病植株,说明新疆野生樱桃李抗南方根结线虫的机制在于侵染后首先引起过氧化氢和超氧阴离子的上升,进而产生HR反应限制了巨细胞的发育,使线虫得不到足够的营养而发育受阻,导致不能形成根结。同时VAD1和PO2基因表达上调可能启动抗病反应,从而使植株产生抗性。     

Genetic diversity analysis in <Emphasis Type="Italic">Plantago ovata</Emphasis> and some of its wild allies using RAPD markers

Plantago ovata, commonly called as the 'desert Indian wheat' is a cultivated and economically important plant of the genus Plantago, a large genus containing ~200 species. It yields Psyllium (Isabgol) which has several health benefits and applications in pharmaceutical, food and cosmetic industries. In view of the genetic uniformity, detection of variability has remained a challenge in this species as the plant lacks inherent variability and has a narrow genetic base. During the present study, Random amplified polymorphic DNA (RAPD) was used to determine genetic relationship and detect whatever little hidden variation exists in this species and some of its wild allies. Limited genetic variability was observed in P. ovata whereas; extensive genetic variability was seen in its wild allies. The genetic distances among different accessions of P. ovata and different species of Plantago, were used to generate a dendrogram.