Development of male-specific markers and identification of sex reversal mutants in papaya

Papaya is a productive and nutritious fruit grown in tropical and sub-tropical regions worldwide. It is polygamous with three sex types: female, male and hermaphrodite. Sex determination in papaya is controlled by an XY sex chromosome system with two slightly different Y chromosomes, Y for males and Y^h for hermaphrodites. Comparative analysis of the hermaphrodite-specific region of Y^h chromosome (HSY) and male-specific region of Y chromosome (MSY) revealed 99.6% sequence identity, which explains why DNA markers that amplify for both males and hermaphrodites have easily been developed, but not for the male trait specifically. We examined the 0.4% sequence differences, and found 1887 indels and 21,088 SNPs between MSY and HSY. The vast majority of indels are single nucleotide or few base pairs. A large male-specific retrotransposon insertion of 8396 bp was used to develop two papaya male-specific markers, PMSM1 and PMSM2 that amplify 585 and 548 bp fragments, respectively. These two markers were tested in 11 gynodioecious and four dioecious varieties along with autosomal DNA marker 71E and male/hermaphrodite marker W11, and the results showed clear separation of male from hermaphrodite and female. PMSM1 and PMSM2 were also used to test the sex type of six sex male-to-hermaphrodite reversal mutants which are crucial materials for validating candidate genes for sex determination in papaya. Our result showed all six mutants were positive for the male-specific markers. These male-specific markers can be used to distinguish gynodioecious and dioecious cultivars in papaya seed market, and facilitate genetic and genomic research for papaya improvement.     

Development and application of a molecular marker for TMV resistance based on <Emphasis Type="Italic">N</Emphasis> gene in tobacco (<Emphasis Type="Italic">Nicotiana tabacum</Emphasis>)

Tobacco mosaic virus (TMV) caused serious loss in yield and quality of tobacco every year. It is a long-term goal to improve the tobacco resistance against TMV by tobacco breeding. N gene was the firstly reported TMV-resistant gene, which showed resistance against all Tobamoviruses except the Ob stain and belonged to the toll-interleukin-1 receptor/nucleotide-binding site/leucine-rich repeat class of plant resistance (R) genes. At present, N gene had already been widely used in tobacco conventional breeding, but there is rare available molecular maker used in marker-assisted selection of TMV resistance. In this study, we designed a pair of primers that specific amplify N gene fragment based on the sequence of N gene intron III, named N-marker. Then, we identified TMV resistance by two selecting methods, PCR with N-marker and inoculated with the TMV-C strain. Results from the two method showed that (1) 13 varieties among 67 tobacco varieties displayed hypersensitive reaction when inoculated with the TMV-C strain, also contained N gene fragments screened by PCR with N-marker; (2) 105 strains of 200 BC1 strains showed resistance against TMV when inoculated with TMV-C strain, meanwhile, 103 of the 105 strains contained N gene fragment verified by PCR with N-marker. Therefore, the N-marker is reliable for high throughput screening of germplasm resources and tobacco breeding materials in selection of N-mediated TMV resistance. Our study not only developed a molecular marker for tobacco breeding, but also identified new germplasm resources that are resistant to TMV.     

甘蓝型油菜烷羟化酶基因MAH1的克隆与表达分析

在植物蜡质合成途径中,中链烷烃羟化酶(mid-chain alkane hydroxylase,MAH)催化烷烃羟基化形成二级醇,进一步氧化为酮。本研究以拟南芥P450依赖性酶CYP96A15/MAH1基因为探针,采用电子克隆与RT-PCR技术,获得2个甘蓝型油菜MAH1的全长编码区序列,分别命名为BnMAH1-1和BnMAH1-2(Gen Bank登录号分别为KT795344和KT795345)。二者ORF长度均为1491 bp,无内含子,核苷酸与氨基酸序列分别有92.4%与90.9%的一致性。根据编码区预测的BnMAH1-1和BnMAH1-2前体蛋白均为包含496个氨基酸残基的多肽链,具有典型的P450蛋白家族保守结构P415x R417x、K螺旋(E359xx R362)、C末端的血红素结合域(F436xx Gx Rx Cx G445)以及氧结合带保守区域(A/G)G309x(D/E)T312(T/S)。NCBI Blast N、氨基酸序列多重比对与系统学分析表明,两者与拟南芥MAH1/CYP96A15同源性最高。实时荧光定量PCR表明,BnMAH1-1与BnMAH1-2主要在甘蓝型油菜茎、叶、花、及角果中表达,其中在叶片中的表达量最高,在根系中的表达量很低,这与角质层蜡质主要沉积在植株地上部分相一致。BnMAH1-1和BnMAH1-2在无蜡粉材料茎、叶片中几乎不表达,表明蜡质的减少与MAH1的转录下调有关。BnMAH1-1与BnMAH1-2受SA、Me JA、ACC、ABA、Na Cl及干旱胁迫诱导表达,其中BnMAH1-1可能在水分胁迫响应中起主要作用。     

新一代温室气体排放情景下安徽省未来气候变化预估分析

基于联合国政府间气候变化专门委员会(IPCC)发布的新一代排放情景,和中国气象局发布的"中国地区气候变化预估数据集V3.0",针对安徽省,笔者选择1971—2000年作为基准期,采用区域气候模式模拟的方法,对比分析安徽省未来气候变化特征,重点评估近期内即未来20年气候变化趋势。模拟结果显示,未来20年不同情景下安徽省平均气温均为上升,全省平均增温幅度约0.9~1.1℃;而降水量变化具有不同特征,较低排放情景下降水量以上升为主,高排放情景则以下降为主。到2050s全省平均气温相比于基准期将升高1.6~1.7℃,升温幅度呈现北高南低的特征;全省降水量相比于基准期将下降50~90 mm,各地降水量均呈下降趋势。另外,通过对降水和气温的模拟,预估未来该地区旱涝演替更加频繁,高温热浪等事件也将进一步频发。     

办公建筑中空调形式对室内外PM2.5浓度相关性的影响

室外PM2.5可通过新风及围护结构缝隙渗透至室内,室外PM2.5较高时尤为明显,结果导致室内空气中的PM2.5浓度上升。为了研究空调形式对室内外PM2.5浓度相关性的影响,在2015年夏季对重庆某办公建筑中采用不同空调形式的室内外PM2.5浓度进行了实测。实测结果发现：集中式空调、分体式空调和非空调房间室内外PM2.5浓度比变化范围分别为0.59～0.76、0.47～0.76、0.71～0.91。室内外PM2.5浓度相关性系数的排序为：集中式空调环境（0.94）> 非空调环境（0.92）> 分体式空调环境（0.77）,研究结果表明,办公建筑的空调形式,对室内外PM2.5浓度的相关性有影响。     

基于数据融合算法的灌区蒸散发空间降尺度研究

采用Landsat和MODIS数据,通过增强自适应融合算法(Enhanced spatial and temporal adaptive reflectance fusion model,ESTARFM)对蒸散发进行空间降尺度,构建田块尺度蒸散发数据集;利用2015年田间水量平衡方法计算的蒸散发数据对融合结果进行评价。在融合蒸散发基础上,结合解放闸灌域2000—2015年间种植结构信息,提取不同作物各自生育期和非生育期内年际蒸散发量,并分析了大型灌区节水改造以来,作物蒸散发占比的年际变化。研究结果表明:融合蒸散发与水量平衡蒸散发变化过程较吻合,小麦耗水峰值出现在6月中下旬—7月初,玉米和向日葵峰值出现在7月份。在相关性分析中,玉米、小麦和向日葵的决定系数R2分别达到了0.85、0.79和0.82;生育期内玉米(5—10月份)、小麦(4—7月份)和向日葵(6—10月份)的均方根误差均不高于0.70 mm/d;平均绝对误差均不高于0.75 mm/d;相对误差均不高于16%。在农田蒸散发总量验证中,融合蒸散发与水量平衡蒸散发相关性较好,两者决定系数达到了0.64。基于ESTARFM融合算法生成的高分辨率蒸散发(ET)结果可靠,具有较好的融合精度。融合结果与Landsat蒸散发的空间分布和差异性一致,7月23日、8月24日和9月1日相关系数分别达到0.85、0.81和0.77;差值均值分别为0.24 mm、0.19 mm和0.22 mm;标准偏差分别为0.81 mm、0.72 mm和0.61 mm。ESTARFM融合算法在农田蒸散发空间降尺度得到较好的应用,可有效区分不同作物蒸散发之间的差异。不同作物在生育期和非生育期内耗水量差别较大;生育期内套种(4—10月份)耗水量最大,达到637 mm,玉米(5—10月份)和向日葵(6—10月份)次之,分别为598 mm和502 mm,小麦(4—7月份)最低为412 mm;非生育期内,小麦(8—10月份)耗水量最大,年均达到214 mm,玉米(4月份)和向日葵(4—5月份)分别为42 mm和128 mm。不同作物多年平均耗水量(4—10月份)差异较小,其年际耗水总量主要随作物种植面积的变化而变化。     

基于茎基部分区边缘拟合的稻株定位方法

准确地定位稻株是水稻株间除草机械作业的前提,提出了侧位俯拍的图像采集方式获取稻株茎基部图像,采用茎基部分区边缘拟合的方法定位稻株,解决了除草期内水稻冠层接连引起的定位不准确问题。首先分析稻株生长形态,探究侧位俯拍稻株茎基部的相机安装参数设定,以及影响成像质量的相关因素,提出了遮光条件下采集图像的方法,构建了具有遮光功能的图像采集系统;其次采用2G-R-B彩色图像灰度化、自动阈值、形态学操作等方法处理并分割稻株图像,检测茎基部边缘并分析其形态特征,并提出了分区边缘拟合定位方法:划分拟合区间为3个子区间,在子区间内进行边缘拟合,以拟合边缘线段中点坐标均值作为子区间茎基部中点,根据中点拟合茎基部中线,以中线中点作为定位的茎基部中心。最后进行了基于茎基部与冠层的稻株定位精度田间对比试验,结果显示:插秧后10~20 d内,基于茎基部的稻株定位误差均在7.0 mm以下,其中10 d、15 d、20 d时的定位误差分别为6.9mm/6.8 mm、5.9 mm/5.8 mm、6.3 mm/6.5 mm(有水条件/无水条件);基于冠层的稻株定位误差在8.0 mm以上,其中插秧20 d后,定位误差超过15 mm以上。试验结果表明,所提出的基于茎基部分区边缘拟合的稻株定位方法,定位精度高,除草适用期长,可满足株间机械除草过程中对稻株精准定位的技术要求。     

液压外骨骼机器人非线性模糊滑模控制方法

针对液压外骨骼机器人系统建模困难的问题,提出一种利用自适应模糊逼近方法来实现对滑模等效控制的逼近,不需要对机器人的未知参数进行预先估计,同时设计可调参数的自适应调节律,增强系统的鲁棒性,引入一种类势能函数设计具有非线性积分项的滑模面,当误差较大时,积分效应适当减弱,防止产生较大的超调量;当误差较小时,积分效应适当增强,减小稳态误差。利用李雅普诺夫方法论证了该闭环控制系统的稳定性,并使用模糊切换方法来消除滑模控制抖振。最后,对液压助力外骨骼机器人系统进行轨迹跟踪及外干扰实验,结果验证了该方法的有效性,控制输出能快速平稳地跟随参考位置信号,且具有一定抗干扰能力。     

河北太行山区典型水土保持林乔木层生物量及碳储量研究

以河北太行山区4种典型水土保持林为研究对象,对混交林(栓皮栎-侧柏)、油松林、栓皮栎林和刺槐林的乔木层各器官生物量、含碳率以及碳储量进行比较研究。结果表明:混交林、油松林、栓皮栎林和刺槐林生物量分别为51.94,86.40,90.19,18.08t/hm^2,栓皮栎林和油松林生物量高于4种水土保持林生物量的均值(61.65t/hm^2),而混交林和刺槐林生物量分别占生物量均值的84.25%,29.33%。不同林分各器官在乔木层生物量中分配顺序均表现为树干>树根>树枝>树叶。4种典型林分各器官含碳率分别为45.16%~58.93%,58.48%~64.61%,51.16%~58.37%,52.35%~62.30%。4种典型林分碳储量为10.10~53.85t/hm^2。不同林分类型各器官碳储量与生物量呈正比关系,与生物量趋势基本相同,碳储量大小表现为油松林>栓皮栎林>混交林>刺槐林。     

Tree-ring response of Larix chinensis on regional climate and sea-surface temperature variations in alpine timberline in the Qinling Mountains

Tree-ring width chronologies of Larix chinensis were developed from the northern and southern slopes of the Qinling Mountains in Shaanxi Province,and climatic factors affecting the tree-ring widths of L.chinensis were examined.Correlation analysis showed that similar correlations between tree-ring width chronologies and climatic factors demonstrated that radial growth responded to climate change on both slopes.The radial growth of L.chinensis was mainly limited by temperature,especially the growing season.In contrast,both chronologies were negatively correlated with precipitation in May of the previous year and April of the current year.Spatial climate-correlation analyses with gridded land-surface climate data revealed that our tree-ring width chronologies contained a strong regional temperature signal over much of northcentral and eastern China.Spatial correlation with seasurface temperature fields highlights the influence of the Pacific Ocean,Indian Ocean,and North Atlantic Ocean.Wavelet coherence analysis indicated the existence of some decadal and interannual cycles in the two tree-ring width chronologies.This may suggest the influences of El Nin˜o-Southern Oscillation and solar activity on tree growth in the Qinling Mountains.These findings will help us understand the growth response of L.chinensis to climate change in the Qinling region,and they provide critical information for future climate reconstructions based on this species in semi-humid regions.