香菇‘申香1644’的选育报告

香菇‘申香1644’是以传统优质栽培品种‘申香215’为亲本,采用多孢自交育种技术选育的新品种。其菌盖纵切面呈凸形,菌盖直径(6.15±0.38) cm,菌盖厚度(2.27±0.42) cm,菇型圆整,菇质紧实,产量高,生物学转化率95%以上。与亲本相比,‘申香1644’在分子标记和栽培性状上均具有明显差异性,其菌盖为浅黄褐色,颜色较亲本浅;菌龄100-105 d,较亲本缩短5-10 d。‘申香1644’菌丝生长适宜温度为22-26 ℃,原基发育适宜温度为16-22 ℃,可在全国范围内进行代料栽培。     

黑木耳‘农黑2号'的选育报告

黑木耳新品种‘农黑2号'是以亲本‘木耳黑龙3号'和‘黑耳9号'的单孢进行杂交,从70个杂交组合中选育的新品种。工厂化栽培条件下,‘农黑2号'单潮出菇鲜重平均为336.4g/袋,干重平均为28.5g/袋,流耳率平均为3.8%。‘农黑2号'具有抗杂能力强、流耳率低、单潮产量高等优点,且能够很好地适应工厂化栽培条件。     

木耳‘农黑1号'的选育报告

黑木耳新品种‘农黑1号'是以黑木耳‘Au5'和‘木耳黑龙3号'作为亲本菌株,通过单孢杂交配对选育而来。工厂化条件下的生产性试验结果表明：‘农黑1号'的第一潮平均鲜耳产量达到322.4g/袋,干耳产量平均为29.0g/袋,出耳率平均为93.3%,生产周期平均为71d。综合来看,‘农黑1号'具有产量高、周期短、出耳率高、一致性好等优点,可作为黑木耳工厂化栽培的优良新品种。     

肺形侧耳‘杭秀2号’的选育

肺形侧耳‘杭秀2号’以‘台秀57’菌株为亲本采用多孢自交育种技术选育而成。该品种菌丝生长适温26-28℃,子实体生长发育适温22-28℃。菌盖扇形,深灰色,菌盖大小平均为4.38cm×3.65cm;菌柄圆柱形,白色,直径平均为1.06cm。鲜菇蛋白质含量2.94%,氨基酸总量2.54%。示范栽培表明,‘杭秀2号’平均生物学效率为71.2%。低温刺激后,经5-6d可完成一茬菇采收。该品种具有出菇整齐、商品性好等优良性状,适于设施栽培。     

斑玉蕈‘闽真5号’的选育报告

斑玉蕈‘闽真5号’品种系以‘闽真3号’与‘白玉-01’作为亲本,通过原生质体单核化杂交育种技术选育获得。该品种菌盖白色,呈半球形,表面斑纹少而小。鲜菇蛋白质含量2.1%,氨基酸总量1.43%。示范栽培表明,‘闽真5号’菌丝最适培养温度20-27 ℃,子实体生长发育温度12-17 ℃,袋栽菌包培养周期为110 d,出菇周期为28 d,平均单袋产量635.17 g/袋。该品种具有栽培周期短、产量高、商品性状好等优良性状,适用于袋栽工厂化周年栽培。     

基于简化基因组测序的大黄鱼耐高温性状全基因组关联分析

利用Illumina HiSeqTM 2500测序平台, 对通过高温胁迫实验筛选得到的20尾耐高温和20尾不耐高温的大黄鱼(Larimichthys crocea)进行了简化基因组测序(SLAF-seq), 每个样本的平均测序深度达到10.26×, 共获得419211个高质量的群体单核苷酸多态性(SNP)位点 。利用TASSEL软件的混合线性模型(MLM)进行全基因组关联分析(GWAS), 共筛选到38个与大黄鱼耐高温性状显著相关的SNP位点(P<2.39E–08)。利用BLAST程序定位每个SNP位点在大黄鱼基因组中的位置, 并分析其周围的功能基因。结果在38个SNPs附近共找到26个已知的功能基因, 这些基因主要与细胞转录、代谢、免疫等功能相关。研究结果可为下一步大黄鱼耐高温分子机制解析及耐高温品种的选育提供参考。     

大口黑鲈转录组SNPs筛选及其与生长的关联分析

为开发人工饲料代替冰鲜杂鱼养殖大口黑鲈的分子标记,以食用冰鲜鱼和配合饲料的同批大口黑鲈为研究材料,利用RNA-Seq（RNA sequencing）技术挖掘SNPs（Single nucleotide polymorphisms）标记,并以关联分析筛选可用于育种的候选标记。转录组进行测序共获得174 M数据,8681个SNPs位点。挑选其中具有表达差异的50个SNPs位点进行SNaPshot分型,结果39个分型成功,其中有4个为假阳性,通过转录组技术开发出SNPs标记35个,成功率为70.0%。为进一步检验这些标记是否可用于评估驯食饲料的大口黑鲈选育研究,研究以327尾摄食人工配合饲料的大口黑鲈为试验材料,SPSS软件进行一般线性模型分析SNPs的不同基因型与生长性状的相关性,结果显示有2个SNPs位点与体质量、全长和体高等生长性状存在显著相关性（P<0.05）,可作为候选标记用于大口黑鲈的分子辅助育种。由于转录组数据直接反应基因的表达情况,从中挖掘与性状相关的优势基因型与分子标记的成功率高,效果较好。同时也为解决大口黑鲈选育研究中标记缺乏提供了有效途径,为选育提供遗传依据、加速育种进程。     

濒危植物羊踯躅全基因组SSR标记开发与鉴定研究

该研究利用基于全基因组限制性酶切位点简化基因组测序技术（RAD seq技术）,开发濒危植物羊踯躅（Rhododendron molle G. Don）全基因组SSR标记,并对3个群体共63份羊踯躅材料进行验证鉴定,为进一步研究羊踯躅的遗传多样性和群体遗传结构以及保护利用提供技术支持。结果显示：（1）羊踯躅基因组测序获得原始数据7.653G bp,过滤后为7.513G bp;经组装发现,羊踯躅171.534 M bp的基因组分布在498 252 contigs中。（2）通过SSR检测,在11 961 SSR位点中获得了11 687对SSR分子标记,并且二核苷酸为基序的重复类型最丰富,达51.76%。（3）随机选取128对SSR标记在6个羊踯躅株系中进行PCR扩增,获得20对高多态性的SSR标记。（4）用所选的20对多态性SSR标记对3个群体共63份羊踯躅材料进行验证分析发现,这些多态性SSR标记位点的等位基因数为4~16个,期望杂合度（H_e）为0.489~0.908。 研究表明,羊踯躅的SSR丰度适中,且二核苷酸为羊踯躅中最丰富的重复序列,该实验进一步证明RAD seq技术是一种经济有效的基因测序方法,实验中开发的SSR引物将有助于进一步研究羊踯躅和其他近缘种的群体结构和多样性。     

10株铜绿假单胞菌的全基因组序列分析

目的 了解不同分离来源铜绿假单胞菌的全基因组基本特征,以此分析基因组多态性及其遗传进化关系。方法 选择10株医源性和食源性来源的铜绿假单胞菌代表性菌株,应用Solexa高通量测序技术对其进行全基因组测序,以此进行多位点序列分型(multilocus sequence typing, MLST),比较各菌株基因组中携带的耐药基因、毒力基因及插入序列(insertion sequence, IS)元件,并通过比较基因组学分析方法拟合泛基因组和核心基因组积累曲线,筛选核心基因SNP构建系统发育分子进化树。结果 10株菌的基因组从6.3～7.0 Mbp大小不一,包含5 868～6 598个基因,平均G+C含量为67.1%;发现10个菌株各具不同的ST型。在这10个菌株的基因组中,共检测到75种耐药基因,包括抗β-内酰胺酶类、抗氨基糖苷类、抗氟喹诺酮类等;共发现188种毒力基因,不同来源菌株间无明显差异;各菌株之间IS元件种类和数量差异较大。分析发现,铜绿假单胞菌具有开放型泛基因组和稳定型核心基因组;10株菌可分为3个进化分支,且不同分离时间和来源无明显相关性。结论 本研究获得10株不同分离来源的铜绿假单胞菌的全基因组序列,初步证实食品及患者分离来源菌株基因组数据无明显相关性,为后续铜绿假单胞菌的分子流行病学和致病性机制研究提供数据参考。     

基于SLAF-seq技术的木薯SNP标记开发

开发大量木薯(Manihot esculenta)特异分子标记可促进木薯种质资源中优良基因的挖掘和利用。本研究以39份木薯种质资源为材料,利用特异性位点扩增片段测序(specific-locus amplified fragment sequencing,SLAF-seq)技术进行测序,结果显示:本次实验双端比对效率为90.83%,酶切效率为91.18%;通过测序获得304.30 Mb的读长数据,各样本读长范围在2 123 094~15 102 046之间,对照数据量最小,仅395 790个读长;样品测序质量值(Q_(30))在95.62%~96.60%之间,均在95%以上;GC含量在41.27%~44.96%之间,平均GC含量为42.89%,对照的GC含量为42.93%。本研究共开发出725 220个SLAF标签,样本的平均测序深度为20.98倍(×),多态性SLAF标签446 789个,共得到2 504 553个群体SNP标记。可见,SLAF-seq技术适用于木薯SNP位点的开发,其效率较高。     

High-density Integrated Linkage Map Based on SSR Markers in Soybean

A well-saturated molecular linkage map is a prerequisite for modern plant breeding. Several genetic maps have been developed for soybean with various types of molecular markers. Simple sequence repeats (SSRs) are single-locus markers with high allelic variation and are widely applicable to different genotypes. We have now mapped 1810 SSR or sequence-tagged site markers in one or more of three recombinant inbred populations of soybean (the US cultivar ‘Jack’ × the Japanese cultivar ‘Fukuyutaka’, the Chinese cultivar ‘Peking’ × the Japanese cultivar ‘Akita’, and the Japanese cultivar ‘Misuzudaizu’ × the Chinese breeding line ‘Moshidou Gong 503’) and have aligned these markers with the 20 consensus linkage groups (LGs). The total length of the integrated linkage map was 2442.9 cM, and the average number of molecular markers was 90.5 (range of 70–114) for the 20 LGs. We examined allelic diversity for 1238 of the SSR markers among 23 soybean cultivars or lines and a wild accession. The number of alleles per locus ranged from 2 to 7, with an average of 2.8. Our high-density linkage map should facilitate ongoing and future genomic research such as analysis of quantitative trait loci and positional cloning in addition to marker-assisted selection in soybean breeding.Key words: EST-derived SSR marker, integrated linkage map, microsatellite marker, polymorphism information content     

中国科学家绘制籼稻高质量参考基因组序列图谱

2005年多国合作的国际水稻(Oryza sativa)基因组测序项目绘制了粳稻(O.sativa subsp.japonica)品种日本晴的参考基因组序列。最近,中国科学家发布了2个籼稻(O.sativa subsp.indica)品种(明恢63和珍汕97)的高质量参考基因组序列,为籼稻的功能基因组学研究和分子育种应用提供了便利。     

The First High-Density Genetic Map Construction in Tree Peony (Paeonia Sect. Moutan) using Genotyping by Specific-Locus Amplified Fragment Sequencing

Genetic linkage maps, permitting the elucidation of genome structure, are one of most powerful genomic tools to accelerate marker-assisted breeding. However, due to a lack of sufficient user-friendly molecular markers, no genetic linkage map has been developed for tree peonies (Paeonia Sect. Moutan), a group of important horticultural plants worldwide. Specific-locus amplified fragment sequencing (SLAF-seq) is a recent molecular marker development technology that enable the large-scale discovery and genotyping of sequence-based marker in genome-wide. In this study, we performed SLAF sequencing of an F1 population, derived from the cross P. ostti ‘FenDanBai’ × P. × suffruticosa ‘HongQiao’, to identify sufficient high-quality markers for the construction of high-density genetic linkage map in tree peonies. After SLAF sequencing, a total of 78 Gb sequencing data and 285,403,225 pair-end reads were generated. We detected 309,198 high-quality SLAFs from these data, of which 85,124 (27.5%) were polymorphic. Subsequently, 3518 of the polymorphic markers, which were successfully encoded in to Mendelian segregation types, and were in conformity with the criteria of high-quality markers, were defined as effective markers and used for genetic linkage mapping. Finally, we constructed an integrated genetic map, which comprised 1189 markers on the five linkage groups, and spanned 920.699 centiMorgans (cM) with an average inter-marker distance of 0.774 cM. There were 1115 ‘SNP-only’ markers, 18 ‘InDel-only’ markers, and 56 ‘SNP&InDel’ markers on the map. Among these markers, 450 (37.85%) showed significant segregation distortion (P < 0.05). In conclusion, this investigation reported the first large-scale marker development and high-density linkage map construction for tree peony. The results of this study will serve as a solid foundation not only for marker-assisted breeding, but also for genome sequence assembly for tree peony.     

猴头菌‘川猴菇1号’的选育

猴头菌新品种‘川猴菇1号’是从黑龙江海林市采集到的野生猴头菌（HT2000）经系统选育而成,适宜四川省凉山州地区及类似生态区域栽培。菌丝生长最适温度为20-25℃,出菇最适温度15-20℃,生育期45-50d,单个子实体130-170g,呈白色至黄白色,中实有弹性,生物转化率85%。     

是金子无论在何处都发光: 玉米和水稻驯化中的趋同选择

野生植物的驯化为人类定居与文明起源奠定了重要基础。在世界范围内不同地区生活的古人类分别对当地不同的野生植物进行了驯化, 而经过驯化的作物常常表现出相似的驯化综合性状。在基因组层面上对趋同选择规律的解析, 可为作物育种提供重要信息与遗传资源。近日, 中国农业大学杨小红/李建生和华中农业大学严建兵领衔的团队从单基因和全基因组2个层次系统解析了玉米(Zea mays)和水稻(Oryza sativa)趋同选择的遗传基础, 发现玉米KRN2与水稻OsKRN2受到了趋同选择, 并通过相似的途径调控玉米与水稻的粒数与产量。他们还发现玉米与水稻在全基因组范围内存在大量趋同选择同源基因对(gene pair), 这些基因在淀粉代谢、糖及辅酶合成等途径特异富集。该研究不仅克隆了在玉米与水稻中均具有重要育种价值的趋同选择同源基因对KRN2/OsKRN2, 而且在全基因组水平上揭示了玉米与水稻趋同选择的规律, 为进一步解析驯化综合性状形成的分子机理及其在育种中的应用奠定了重要理论基础。     

毛木耳‘昊阳黄背2号’的选育报告

毛木耳新品种‘昊阳黄背2号’是以采自什邡市龙泉山的野生毛木耳子实体经系统选育而成,适宜四川省成都市、德阳市等种植毛木耳的区域栽培。菌丝生长最适温度为24-30 ℃,子实体最适生长温度为25-28 ℃。耳片呈耳形或不规则形,子实体丛生;耳片边缘光滑,颜色深红褐色。平均每袋干耳产量为203 g。     

基于报告基因敲入构建果生刺盘孢细胞核荧光标记菌株

运用基因敲入技术,将GFP、mCherry整合到果生刺盘孢 Colletotrichum fructicola组蛋白histone H1位点,实现融合表达,获得细胞核荧光标记菌株。基于标记菌株可以对分生孢子、营养菌丝、附着胞、侵染菌丝等结构中的细胞核进行实时活体观察。果生刺盘孢存在性亲和的“+”、“-”型菌株分化,GFP“+”型菌株和mCherry“-”型菌株接触形成明显杂交线,杂交线上单子囊内含红绿两种孢子,表明“+”、“-”型菌株间发生杂交;杂交线上子囊壳壁表达mCherry,表明由“-”型菌株发育而来。本研究构建的核荧光标记菌株将是研究果生炭疽菌细胞周期调控和有性繁殖过程的重要材料。     

Genome-Wide Identification of SSR and SNP Markers Based on Whole-Genome Re-Sequencing of a Thailand Wild Sacred Lotus (Nelumbo nucifera)

Genomic resources such as single nucleotide polymorphism (SNPs), insertions and deletions (InDels) and SSRs (simple sequence repeats) are essential for crop improvement and better utilization in genetic breeding. However, the resources for the sacred lotus (Nelumbo nucifera Gaertn.) are still limited. In the present study, to dissect large-scale genomic molecular marker resources for sacred lotus, we re-sequenced a Thailand sacred lotus cultivar ‘Chiang Mai wild lotus’ and compared with the reported lotus genome ‘Middle lake wild lotus’. A total of 3,180,059 SNPs, 328, 251 InDels and 14,191 SVs were found between the two genomes. The functional impact analyses of these SNPs indicated that they may be involved in metabolic processes, binding, catalytic activity, etc. Mining the genome sequences for SSRs showed that 191,657 SSRs were identified with a frequency of one SSR per 4.23 kb and 103,656 SSR primer pairs were designed. Furthermore, 14, 502 EST-SSRs were also indentified using the available RNA-seq data in the NCBI. A subset of 150 SSRs (genomic and EST-SSRs) was randomly selected for validation and genetic diversity analysis. The genotypes could be easily distinguished using these SSR markers and the ‘Chiang Mai wild lotus’ was obviously differentiated from the other Chinese accessions. This study provides considerable amounts of genomic resources and markers for the quantitative trait locus (QTL) identification and molecular selection of the species, which could have a potential role in various applications in sacred lotus breeding.