中国樱桃InDel标记开发及其在蔷薇科果树中通用性评价

对栽培和野生中国樱桃进行De novo基因组测序和比对，在全基因组范围内检测筛选出50 617个InDel位点，并挑选均匀分布的200个标记进行验证，其中，27个标记在中国樱桃种内表现出多态性。利用多态性标记，对不同来源的192份中国樱桃（168份栽培和24份野生）种质进行基因型分型，共检测到60个等位基因（N_a），平均每个标记2.2个；基因多样性指数（H_s）为0.010 ~ 0.500，平均0.239；多态性信息含量（PIC）为0.010 ~ 0.375，平均0.198，表明中国樱桃遗传基础相对狭窄。根据聚类结果和地理分布将192份中国樱桃种质大致划分为5个群体。筛选出在蔷薇科李亚科樱属、李属、桃属、杏属、苹果亚科梨属、苹果属和蔷薇亚科草莓属、悬钩子属等重要果树共8个属156个个体中表现出较好通用性标记34个。物种与中国樱桃亲缘关系越近，InDel标记的多态性越高。     

大花君子兰叶绿体基因组及其特征

采用Illumina MiSeq测序平台对大花君子兰（Clivia miniata）叶片总DNA进行测序，通过组装获得了其叶绿体基因组（cpDNA）全长序列（158 114 bp）。对其cpDNA注释得到135个基因，包含87个蛋白编码基因、40个tRNA基因和8个rRNA基因。采用生物信息学方法对获得的cpDNA进行简单序列重复（SSR）分析和密码子偏好性分析。结果显示：①大花君子兰cpDNA中共有61个SSR位点，其中单核苷酸、二核苷酸、三核苷酸、四核苷酸、五核苷酸和六核苷酸重复数分别为38、9、2、8、3和1个，多数SSR分布在基因间隔区；②大花君子兰cpDNA密码子偏爱以A或U（T）结尾，亮氨酸使用频率最高，半胱氨酸使用频率最低。基于24种植物的cpDNA全长和23种植物的叶绿体ycf2基因序列进行系统发育分析，结果显示大花君子兰与石蒜科植物在同一分支，显示最近的亲缘关系，支持大花君子兰属于石蒜科。基于叶绿体ycf2的系统发育分析结果与基于cpDNA全长的系统发育分析研究结果大部分相同，支持ycf2基因可以代替cpDNA全长用于植物系统发育分析。     

番茄斑萎病毒病抗性种质筛选及人工接种鉴定方法优化

对154份番茄材料(包括普通番茄76份,樱桃番茄78份)进行两年两次田间番茄斑萎病毒病的病情指数调查,筛选出14份对番茄斑萎病毒病具有稳定抗性的番茄材料,利用sw-5-2共显性SCAR标记对田间表现抗病的材料进行分子标记鉴定,发现3份抗病材料携带抗番茄斑萎病毒病的Sw-5基因。为缩短番茄斑萎病毒病人工接种鉴定周期,以含有sw-5的抗病材料H8和感病材料M82为研究对象,设置4、6、8、10片真叶4个接种时期,分别在接种后14、21、28 d进行病情指数调查和抗性分级,结果表明,6片真叶期接种,接种后28 d进行病情调查即可有效鉴别植株番茄斑萎病毒病抗性,与8、10片真叶期接种效果相同,人工接种抗病性鉴定效率显著提升。     

胡葱黄条病毒吉林毛葱分离物全基因组序列测定与分析

使用RT-PCR方法从吉林省疑似感染胡葱黄条病毒(Shallot yellow stripe virus,SYSV)的分蘖洋葱(毛葱)叶片中获得了SYSV吉林毛葱分离物SYSV-JL(MN607702)的全基因组序列。SYSV-JL的全长序列为10?427 nt,与GenBank已登录的3条SYSV全长或近全长序列在多个基因上保持着较高的序列一致性,但在P1基因上核苷酸和氨基酸序列一致性较低,分别为69.5%～84.2%和60.9%～76.9%,进一步通过变异分析发现,P1基因包含369个变异位点,表现出较为明显的高变异特征。系统发育分析显示,不同SYSV分离物具有较为明显的地区差异,SYSV-JL分离物与多个中国分离物系统发育关系较近。     

‘鸭梨’、‘新高’果实发育过程中果肉及种子有机酸含量的变化

为了研究梨种子对果实生长发育的影响,采用高效液相色谱(HPLC)技术对‘鸭梨’和‘新高’果实发育过程中果肉和种子中的有机酸含量进行了分析。结果表明：两个品种果肉中主要的有机酸组分均为苹果酸和奎尼酸,发育初期‘鸭梨’与‘新高’果肉中各组分的比率相近,但果实成熟时‘新高’果肉中苹果酸的比率59.98%显著高于‘鸭梨’的37.51%；两个品种种子中的有机酸组分比率的动态变化相近,与果肉相反,主要表现在奎尼酸比率的增加和苹果酸比率的减少。两个品种果肉和种子中的总酸、草酸、奎尼酸,苹果酸,莽草酸,柠檬酸含量均随着果实的成熟呈下降趋势,‘鸭梨’果肉中的奎尼酸、莽草酸、柠檬酸含量高于‘新高’；‘新高’在果实成熟时种子中的总酸含量4.59mg/g显著低于‘鸭梨’13.00mg/g,‘鸭梨’成熟果实种子中的草酸、奎尼酸、莽草酸含量显著高于‘新高’；在果实发育初期,种子中的苹果酸含量极显著高于果肉,在果实成熟时则显著低于果肉。     

Simulation modelling of potato virus Y spread in relation to initial inoculum and vector activity

Potato virus Y(PVY) is a non-persistent virus that is transmitted by many aphid species and causes significant damage to potato production. We constructed a spatially-explicit model simulating PVY spread in a potato field and used it to investigate possible effects of transmission efficiency, initial inoculum levels, vector behavior, vector abundance, and timing of peak vector activity on PVY incidence at the end of a simulated growing season. Lower PVY incidence in planted seed resulted in lower virus infection at the end of the season. However, when populations of efficient PVY vectors were high, significant PVY spread occurred even when initial virus inoculum was low. Non-colonizing aphids were more important for PVY spread compared to colonizing aphids, particularly at high densities. An early-season peak in the numbers of noncolonizing aphids resulted in the highest number of infected plants in the end of the season, while mid-and late-season peaks caused relatively little virus spread. Our results highlight the importance of integrating different techniques to prevent the number of PVY-infected plants from exceeding economically acceptable levels instead of trying to control aphids within potato fields. Such management plans should be implemented very early in a growing season.     

利用CRISPR/Ca9技术靶向编辑芥蓝BoaZDS

以芥蓝(Brassicaoleraceavar.alboglabra)为材料,以ζ–胡萝卜素脱氢酶(ζ-Carotene desaturase,ZDS)基因为目标基因,建立其CRISPR/Cas9基因组编辑体系。在BoaZDS的编码区近5′端选择靶位点,构建了CRISPR/Cas9表达载体,通过农杆菌介导的遗传转化方法获得了19个芥蓝转基因阳性植株,Sanger测序分析发现其中13株成功突变,CRISPR/Cas9载体在芥蓝上的突变效率为68.42%,且所有突变植株均表现出明显的白化表型。     

稳定同位素技术鉴别库尔勒香梨产地可行性研究

为开展碳、氮、氢、氧(C、H、O、N)稳定同位素鉴别库尔勒香梨产地的可行性研究,明确新疆库尔勒香梨δ¹³C、δ¹⁵N、δ²H、δ¹⁸O值,本研究以甘肃香梨和陕西红香酥梨作为对照,采用稳定同位素比值质谱仪(IRMS)分析新疆库尔勒香梨与对照样品的δ¹³C、δ¹⁵N、δ²H、δ¹⁸O值,并采用正交校正的偏最小二乘判别分析(OPLS-DA) 鉴别新疆库尔勒香梨产地真实性。结果表明,新疆、甘肃两个产区香梨与陕西产区红香酥梨的δ¹³C、δ¹⁵N、δ²H、δ¹⁸O值存在显著差异(P<0.05)。新疆产区的66份库尔勒香梨判别准确率为95.45%;甘肃产区的20份香梨样品判别准确率为60%;陕西产区的23份红香酥梨样品判别准确率为95.65%。C、N、H、O 4种稳定同位素可有效鉴别新疆库尔勒香梨和陕西红香酥梨,但在新疆库尔勒香梨与甘肃香梨鉴别方面,误判率较高,如果能结合其他鉴别技术,则有可能出现较好的鉴别效果。本研究为库尔勒香梨产地真实性鉴别提供了科学依据。     

Panax notoginseng saponins suppress oxygen-glucose deprivation/reoxygenation-induced pyroptosis of SH-SY5Y cells

AIM To investigate the effect of Panax notoginseng saponins (PNS) on pyroptosis of SH-SY5Y cells induced by oxygen-glucose deprivation/reoxygenation (OGD/R). METHODS The OGD/R was conducted to induce ischemia/reperfusion injury in SH-SY5Y cells. The effects of PNS on the viability (detected by CCK-8 assay) and membrane permeability [indicated by lactate dehydrogenase (LDH) leakage and propidium iodide (PI) staining positive cell proportion] of OGD/R-induced SH-SY5Y cells were observed. The protein levels of gasdermin D (GSDMD), GSDMD N-terminal fragment (GSDMD-N), caspase-1 and caspase-4, and the release of interleukin-1β (IL-1β) and IL-18 in the cells were also determined. RESULTS After exposure to OGD/R, the viability of SH-SY5Y cells dramatically decreased (P<0.01), while the LDH leakage, the PI staining positive cell proportion, the protein levels of GSDMD, GSDMD-N, caspase-1 and caspase-4, and the release of IL-1β and IL-18 were significantly increased (P<0.01). However, PNS treatment enhanced the viability of SH-SY5Y cells inhibited by OGD/R (P<0.01), but reduced the leakage of LDH and the percentage of PI staining positive cells (P<0.05 or P<0.01). Moreover, PNS reversed the increases in the protein levels of GSDMD, GSDMD-N, caspase-1 and caspase-4 and the release of IL-1β and IL-18 in OGD/R-induced SH-SY5Y cells (P<0.05 or P<0.01). CONCLUSION Treatment with PNS alleviates OGD/R-induced injury in SH-SY5Y cells. Its mechanism may be related to inhibition of SH-SY5Y cell pyroptosis induced by OGD/R.