卷丹转基因体系构建及岷江百合LrCCoAOMT的导入

以卷丹（Lilium lancifolium Thunb.）无菌小鳞片为试材,通过切片处理、优化农杆菌侵染浓度与时间以及重悬液和共培养基成分,构建农杆菌介导的高效遗传转化体系。结果表明,MS + 1.5 mg ? L-1 6-BA + 0.5 mg ? L-1 NAA + 30 g ? L-1蔗糖是切片芽分化的最佳培养基,添加100 mg ? L-1抗坏血酸能有效抑制褐化并促进不定芽增殖。MS + 2.0 mg ? L-1 NAA + 30 g ? L-1蔗糖是生根诱导的最佳培养基。抗生素敏感性测试发现,培养基中添加Kan 100 mg ? L-1或Hyg 75 mg ? L-1 结合Cef 400 mg ? L-1适宜抗性筛选。GUS染色分析表明,以去除大量元素的改良MS + 100 μmol ? L-1 AS和去除大量元素的改良MS + 1.0 mg ? L-1 6-BA + 1.0 mg ? L-1 NAA + 100 μmol ? L-1 AS为重悬液和共培养基,将切片在农杆菌菌液浓度OD600为0.4,侵染15 min,获得81.72% 瞬时转化率和25.2% 稳定遗传转化率。将岷江百合LrCCoAOMT转化卷丹,分子检测和GUS染色分析表明已获得转基因阳性株系。     

基于叶绿体DNA分析的楸子种质遗传多样性研究

利用4对叶绿体DNA引物扩增49份楸子[Malus prunifolia（Willd.）Borkh.]种质资源的4个叶绿体DNA基因间区trnH-psbA、trnS-trnG spacer + intron、trnT-5′trnL和5′trnL-trnF序列,基于4个叶绿体DNA基因间区的序列变异,从母系遗传的角度评价楸子的遗传多样性水平。结果显示：4个叶绿体DNA基因间区序列经测序、拼接、比对和合并之后的片段长度为3 790 bp,共有173个多态性变异位点,其中包含2个单一突变位点、20个简约信息位点和151个插入/缺失位点。在49份楸子种质中,trnH-psbA、trnS-trnG spacer + intron、trnT-5′trnL和5′trnL-trnF区域的变异位点的数量分别为26个、25个、120个和2个,单倍型数量分别为9个、7个、8个和3个,合并之后的叶绿体DNA片段的单倍型有14个。核苷酸多样性和单倍型多样性最高的区域均为trnH-psbA（Hd = 0.775,Pi = 0.02143）,最低的为5′trnL-trnF（Hd = 0.481,Pi = 0.00072）。49份楸子种质4个叶绿体DNA区域合并后的遗传多样性较高（Hd = 0.854,Pi = 0.00949）。Tajima’s D检验中,4个叶绿体DNA区域在P > 0.10水平上均不显著,楸子的4个叶绿体DNA区域在进化上遵循中性进化模型。楸子的遗传变异主要存在于群体内部,不同居群间基因交流频繁,多数居群间遗传分化较少,与地理距离不完全相关。     

甜瓜嫩果皮颜色遗传规律研究

为探究甜瓜嫩果皮颜色的遗传规律,以甜瓜嫩果皮颜色为深绿色的S26和青色的S28为亲本材料,通过构建BC1群体,利用卡方测验,分析回交群体BC1的深绿色嫩果皮和青色嫩果皮的分离比例,开展甜瓜嫩果皮的遗传规律分析,以对嫩果皮颜色基因进行初步定位。结果表明：BC1群体表型表现为深绿色和青色2种,且群体比例为1∶1,从而确定颜色性状为单基因控制且深绿色为显性性状。通过分离群体分组分析法（BSA）和混池测序结果可以看出,控制甜瓜嫩果皮颜色的基因位于chr04的端部位置,这为后续基因的精细定位以及克隆提供了研究基础。     

Strategy to identify resistance to white mould associated with high yield for irrigated common bean in Brazil

Development of common bean cultivars with partial white mould resistance through breeding techniques has been a challenge in Brazil. As yet, lines/cultivars from breeding programmes have not been investigated for resistance; therefore, this study screened 107 lines/cultivars for their reactions to white mould in 14 preliminary trials conducted under irrigation. Thirteen resistant lines/cultivars (three of Andean origin) and six Mesoamerican cultivars (three intermediately resistant and three susceptible) were selected for further investigation. These lines/cultivars and the resistant control A195 were evaluated in six advanced trials and two straw tests to assess the effectiveness of the screening procedure. In 11 preliminary trials, screenings were performed under moderate/high or higher disease pressure. These pressures occurred in two advanced trials in which, when yields were averaged across moderate/high and high pressures, 10 Mesoamerican lines/cultivars selected for resistance yielded 14%, 23%, and 38% more than intermediately resistant cultivars, A195, and susceptible cultivars, with median disease ratings (1–9 scale) of 4.5, 5.7, 5.7, and 6.7, respectively. In the straw test, three Andean lines/cultivars (A195 included) and two susceptible cultivars in the field were among those with the highest levels of physiological resistance. Thus, field rating under high disease pressure and greenhouse rating did not correlate significantly, suggesting that field trials are critical to evaluating resistance and to identifying high-yielding beans. Therefore, lines/cultivars from breeding programmes assessed in field trials may provide a low cost and fast way to identify high-yielding bean cultivars with partial resistance to white mould in the subtropical southern hemisphere.     

Salinity tolerance of wild barley Hordeum vulgare ssp. spontaneum

Salinity tolerance of 47 wild barley genotypes and six barley cultivars was evaluated under control and salinity stress (300 mM NaCl) conditions. Shoot and root dry weight (DW), plant height, membrane stability index (MSI), relative water content, survival rate, leaf malondialdehyde (MDA) and proline contents, root and leaf Na, K, Ca and K/Na ratio, and chlorophyll a fluorescence were measured. Salinity stress caused significant increase in the MDA, proline content, Na and Ca concentrations of the roots and leaves, but resulted in a decrease in the other traits. H. spontaneum genotypes were considerably less affected by the salinity than the genotypes of H. vulgare. Plant survivability was negatively correlated with the Na concentration (r =−.66) but positively correlated with the leaf K/Na ratio (r = .67) and MSI (r = .68). Tolerance mechanisms such as ion exclusion (Na) were likely to be present in the wild barley causing K/Na homeostasis as well as the much lower root and shoot Na, resulting in the higher survival rate.     

川麦42和川农16抗穗发芽QTL定位及聚合效应分析

【目的】小麦穗发芽严重影响小麦产量和品质,是全球小麦生产面临的重大问题之一。通过鉴定挖掘抗穗发芽QTL,聚合穗发芽抗性位点,选育抗穗发芽小麦品种,为四川小麦穗发芽抗性改良提供技术和材料支撑。【方法】以川麦42/川农16重组自交系（RIL,F₈）为材料,于2016—2018年分别在2个环境下对RIL群体进行籽粒发芽指数（GI,2016和2018）、籽粒发芽率（GR,2016和2018）和整穗发芽率（SGR,2017和2018）3个穗发芽指标测定。利用90K SNP芯片构建的遗传图谱检测全基因组穗发芽相关QTL,并分析抗性QTL聚合效应。【结果】双亲间GI、GR和SGR指标值差异显著,亲本川农16穗发芽抗性明显优于亲本川麦42。共检测到11个与穗发芽抗性有关的QTL,主要分布在2B、2D、3A、3D、4A、5A、5B和6B染色体上。5B染色体上检测到的单个环境表达的整穗发芽QTL解释的表型变异率最大,达到29%;在2D和3A染色体上检测到的整穗发芽主效QTL,以及5A染色体上检测到的与种子休眠相关的籽粒发芽主效QTL,在2个环境下均能表达,其抗穗发芽等位变异均来源于川农16。基因型分析发现,RIL群体中不同株系聚合抗性QTL的数量变幅为1—9个,表现为抗穗发芽的株系均携带4—9个与穗发芽相关的抗性QTL。重组自交系群体中6个株系GI、GR和SGR值均在15%以下,表现出高抗穗发芽特性;这6个优异株系聚合了多个与穗发芽相关的抗性QTL,且均聚合了川麦42在4A染色体上的微效QTL（QGi.saas-4A和QGr.saas-4A）,以及川农16在2D和5B染色体上的主效QTL（QSgr.saas-2D和QSgr.saas-5B）;编号为104和125的优异株系已通过审定,定名为川麦104和川麦64。其中,川麦104于2012年同时通过国家和四川省审定,其抗穗发芽能力强,产量、品质、抗病等优良性状突出,聚合了7个正向穗发芽QTL,包括2B、2D和5B染色体上来源于川农16的4个抗性QTL（QGi.saas-2B、QGr.saas-2B、QSgr.saas-2D和QSgr.saas-5B）,以及4A和6B染色体上来源于川麦42的3个QTL（QGi.saas-4A、QGr.saas-4A和QGr.saas-6B）;近年来,川麦104已成为西南麦区小麦育种的核心亲本,育成小麦品种（系）18个。【结论】共检测到11个抗穗发芽QTL,其中3个来源于川麦42,8个来源于川农16;RIL群体中的抗穗发芽株系均携带4—9个抗性QTL,优异株系川麦104和川麦64高抗穗发芽,均聚合了7个穗发芽抗性QTL。     

An efficient method to produce segregating populations in quinoa (Chenopodium quinoa)

Quinoa offers a promising alternative for staple food, considering its outstanding nutritional value and tolerance to abiotic stresses. To develop breeding programmes in quinoa, a reliable crossing method for increasing the genetic variation is required. In the following study, we aimed to develop segregating populations in quinoa. We tested the efficiency of three different crossing methods (hand emasculation, warm water emasculation and no emasculation). Moreover we developed a two-stage selection strategy based on morphological traits and molecular markers for the selection of hybrid plants. We reported hand emasculation to be the most efficient crossing method, followed by warm water emasculation and no emasculation. Our results demonstrated that crosses in quinoa can be successfully performed, despite its complicated flower structure and high self-pollination rate. Additionally, we developed 30 segregating populations from crosses between accessions of different origins with varying phylogenetic relationship, which offers a promising perspective for quinoa breeding programmes in the future.     

Genetic variation of seedling traits responded to brassinosteroid and gibberellin inhibitors in maize (Zea mays) doubled haploid lines

The purpose of this study was to examine the genotypic variation in maize doubled haploid (DH) lines response to brassinosteroid and gibberellin inhibitors. Plant responses to hormone inhibitors were determined in growth chamber experiments using germination paper for three different seedling treatments: application of propiconazole (Pcz), uniconazole (Ucz) or water (control). Mesocotyl length (ML) was more sensitive to hormone inhibitors, especially to the Ucz treatment, than other seedling traits. ML was significantly correlated with other traits in the Ucz treatment. All the seedling traits showed moderate-to-high broad sense heritability values, ranging from 0.39 to 0.82. The Euclidian genetic distances of inbred line pairs ranged from 1.27 to 19.94, indicating there was a high level of variability across the maize DH lines used in this study. DH lines with extreme MLs were identified, which can provide valuable breeding resources for improving abiotic stress tolerance, and for further genetic studies.     

湖南省饭豆地方种质资源遗传多样性的RAPD和ISSR分析

为揭示湖南省饭豆地方品种的遗传多样性,促进种质资源的有效保护和利用。本研究利用RAPD和ISSR标记分析了源于湖南省各地36份古老饭豆地方品种的遗传多样性,并采用UPGMA方法进行了遗传聚类分析。结果表明：12个RAPD引物和4个ISSR引物在供试材料中共扩增产生81条带型清晰的条带,平均每个引物扩增产生5.06个条带;其中59个为多态性条带,占总扩增条带的72.84%,具有多态性引物的PIC值在0.397~0.968之间;通过UPGMA法进行聚类分析表明供试材料的遗传相似系数在0.605~0.877之间,供试材料被分为4大类,且聚类结果表现明显的地域特征。本研究结果可为本省饭豆种质资源保护和育种研究提供理论依据。