Generating Marker-Free Transgenic Tobacco Plants by Agrobacteriummediated Transformation with Double T-DNA Binary Vector

We have developed a 'double T-DNA' binary vector system for generating selectable marker-free transgenic plants by Agrobacterium-mediated transformation. The 'double T-DNA' binary vector pDLBRBbarm which carried two independent T-DNAs, one containing a selectable marker neomycin phosphotransferase (nptII) gene and the other a bargene, was constructed. Transgenic tobacco (Nicotiana tabacum L.) plants were then produced by Agrobacterium-mediated transformation with this vector. Frequency of the primary transformants co-integrated with npt II gene and bar gene was 59.2%. Segregation of two T-DNA regions was found in 3 out of 4 T-1 lines from co-transformed T-0 plants with nptII and bar PPT-resistant and kanamycin-sensitive plants were in approximate 19.5% of the T-1 plants. The result indicated that this 'double T-DNA' vector system could be a workable approach to generate transgenic plants free from selectable marker genes. Co-transformation of nptII gene and bar gene to plants with mixtures of Agrobacterium tumefaciens strains containing single T-DNA vectors was also tested. Frequency of co-transformed plants was 20.0%-47.7% and relatively low as compared with that of 'double T-DNA' vector system.

Correlation Analysis Between Yield Components and Yield Per Plant of Double-low Hybrid Rapeseed

[Objective] The experiment aimed to study the relation between yield components and yield per plant of double-low hybrid rapeseed and provide reference for breaking yield limitation of rapeseed and culturing new double-low hybrid rapeseed variety. [Method]The yield components and yield per plant of two cross combination of double-low hybrid rapeseed (B02, D04) and Shuza 6 were correlatively analyzed and compared, besides, the path analysis was also carried on to them. [Result] Among B02, D04 and Shuza No.6, effective pod number per plant and seeds per silique, seeds per pod and 1 000-grain weight were all negative correlation. In high yield hybrid, pod number per plant, seeds per pod had more impaction on yield per plant than 1 000-grain weight and the difference was at 0.01 significant level. In the control variety Shuza No.6, the impactions of pod number per plant and seeds per pod on yield per plant were bigger than that of 1 000-grain weight on yield per plant, however, the difference was not significant. [Conclusion] The improvement of effective pod number per plant was an important aim of breeding work of double low rapeseed breeding in Sichuan ecological region.

Mapping QTLs with epistatic effects and QTL×environment interactions for plant height using a doubled haploid population in cultivated wheat

Quantitative trait loci （QTLs） for plant height in wheat （Triticum aestivum L.） were studied using a set of 168 doubled haploid （DH） lines, which were derived from the cross Huapei 3/Yumai 57. A genetic linkage map was constructed using 283 SSR and 22 EST-SSR markers. The DH population and the parents were evaluated for wheat plant height in 2005 and 2006 in Tai＇an and 2006 in Suzhou. QTL analyses were performed using the software of QTLNetwork version 2.0 based on the mixed linear model. Four additive QTLs and five pairs of epistatic effects were detected, which were distributed on chromosomes 3A, 4B, 4D, 5A, 6A, 7B, and 7D. Among them, three additive QTLs and three pairs of epistatic QTLs showed QTL×environment interactions （QEs）. Two major QTLs, Qph4B and Qph4D, which accounted for 14.51% and 20.22% of the phenotypic variation, were located similar to the reported locations of the dwarfing genes Rhtl and Rht2, respectively. The Qph3A-2 with additive effect was not reported in previous linkage mapping studies. The total QTL effects detected for the plant height explained 85.04% of the phenotypic variation, with additive effects 46.07%, epistatic effects 19.89%, and QEs 19.09%. The results showed that both additive effects and epistatic effects were important genetic bases of wheat plant height, which were subjected to environmental modifications, and caused dramatic changes in phenotypic effects. The information obtained in this study will be useful for manipulating the QTLs for wheat plant height by molecular marker-assisted selection （MAS）.

Stem flow of seed-maize under alternate furrow irrigation and double-row ridge planting in an arid region of Northwest China

Maize is widely planted throughout the world and has the highest yield of all the cereal crops. The arid region of North- west China has become the largest base for seed-maize production, but water shortage is the bottleneck for its long-term sustainability. Investigating the transpiration of seed-maize plants will offer valuable information for suitable planting and irrigation strategies in this arid area. In this study, stem flow was measured using a heat balance method under alternate furrow irrigation and double-row ridge planting. Meteorological factors, soil water content （e）, soil temperature （Ts） and leaf area （LA） were also monitored during 2012 and 2013. The diurnal stem flow and seasonal dynamics of maize plants in the zones of south side female parent （SFP）, north side female parent （NFP） and male parent （MP） were investigated. The order of stem flow rate was： SFP〉MP〉NFP. The relationships between stem flow and influential factors during three growth stages at different time scales were analyzed. On an hourly scale, solar radiation （Rs） was the main driving factor of stem flow. The influence of air temperature （Ta） during the maturity stage was significantly higher than in other periods. On a daily scale, Rs was the main driving factor of stem flow during the heading stage. During the filling growth stage, the main driving factor of NFP and MP stem flow was RH and Ts, respectively. However, during the maturity stage, the environ- mental factors had no significant influence on seed-maize stem flow. For different seed-maize plants, the main influential factors were different in each of the three growing seasons. Therefore, we identified them to accurately model the FP and MP stem flow and applied precision irrigation under alternate partial root-zone furrow irrigation to analyze major factors affecting stem flow in different scales.

Formula for Determining Number of Basic Seedlings at Scattered-Planting with Seedling Dry-Raised on Plastic Trays in Double-Season Rice

The tiller emergence in seedling nursery beds and field, and panicle formation in the field were investigated under scattered-planting with seedling dry-raised on plastic trays in double-season rice. A significant difference was noted in the non-synchronously-emerged tillers （the tillers that formed from latent buds and did not emerge following the normal tillering law on seedling nursery beds and recovered to grow after scattered-planting or transplanting） as well as the percentage of the available synchronously-emerged tillers between seedlings raised on plastic trays under dry-land conditions （DPT） and seedlings raised on nursery beds under wetland conditions （WB）. The seedlings under DPT had some non-synchronously-emerged tillers, but those under WB had not. Therefore, the traditional formula for determining the number of rice seedlings was improved, and the formula for determining the number of basic seedlings under scattered planting with DPT in double-season rice was introduced. For early rice, it was X=Y/{（I＋t1r1）[1＋（N-n-SN）Rr2]＋（SN-3-t1）R2r5}, and for late rice, it was X=Y/{（1＋t1r1）[1＋（N-n-SN）Rr2]＋（N-n-SN-3）Rr2R1r3＋（SN-3-t1）R2r5}. Where, X represents reasonable number of basic seedlings per unit area at scattered-planting; Y, number of fitting panicles per unit area; t1, total number of tillers per plant; r1, percentage of the total available tillers; N, total number of leaves of the main culm; n, total number of elongated internodes in the main culm; SN, seedling leaf ages at scattered-planting; R, percentage of the primary tillers emerged in available node-position; r2, percentage of the available primary tillers; R1, percentage of the secondary tillers in the field （except the secondary tillers of the seedlings）; r3, percentage of the available secondary tillers; R2, percentage of the asynchronously-emerged tillers after scattered-planting; r5, percentage of the available non-synchronously-emerged tillers after scattered-planting.

In vitro Regeneration of First-generation Inbred Progenies of Cucurbita pepo L. Double Haploids

Using seeds of inbred progenies of Cucurbita pepo L. DH12-11409 double haplolds as experimental materials, the effects of different hormone combinations, explant types and seedling ages on adventitious bud regeneration and rooting of C. pepo L. were investigated. According to the results, inoculating cotyledonary nodes of yellow-green cotyledons from 5-d-old C. pepo L. double haploids to MS ＋ 30 g/L Suc ＋ 8 g/L Agar ＋ 2.0 mg/L 6-BA ＋ 0.1 mg/L NAA led to the best results with the induction frequency of 90.0% and differentiation coefficient of 8.5. MS medium with addition of 0.05 or 0.1 mg/L NAA led to the highest rooting rate. Regenerated seedlings with 5 - 6 true leaves exhibited the highest survival rate of 90.0%, which was the optimal period for domestication and transplanting of regenerated seedlings. This study laid a solid foundation for high-efficiency utilization of heterosis of C. pepo L.

Construction of Double Right-Border Binary Vector Carrying Non-Host Gene Rxo1 Resistant to Bacterial Leaf Streak of Rice

Rxol cloned from maize is a non-host gene resistant to bacterial leaf streak of rice. pCAMBIA1305-1 with Rxo1 was digested with Sca I and NgoM IV and the double right-border binary vector pMNDRBBin6 was digested with Hpa I and Xma I. pMNDRBBin6 carrying the gene Rxo1 was acquired by ligation of blunt-end and cohesive end. The results of PCR, restriction enzyme analysis and sequencing indicated that the Rxo1 gene had been cloned into pMNDRBBin6. This double right-border binary vector, named as pMNDRBBin6-Rxol, will play a role in breeding marker-free plants resistant to bacterial leaf streak of rice by genetic transformation.

旨在培养科研素养的理工科研究生课程教学改革——以河南大学高级植物生理学为例

培养科研创新能力是研究生教育的核心任务之一,也是研究生培养质量的集中体现。以植物逆境生物学为特色的河南大学生物学科入选国家“双一流”建设行列,对研究生科研创新能力培养提出新的更高要求。高级植物生理学是植物逆境生物学的核心课程,该课程围绕研究生科研素养提升的核心目标采取一系列教学改革举措,通过科研反哺教学、打造精良的教学团队、更新和拓展教学内容、改革教学手段和教学方式、融入课程思政,以及双语教学和考核方式等方面的改革实践,使科研素养培养始终贯穿于课程教学的全过程中,为提升研究生创新意识和创新能力筑牢基础。此课程的教学改革实践为“双一流”大学理工科研究生课程建设提供新的借鉴。

“双季同种”水稻新品种松雅7号的选育与思考

“双季同种”是笔者根据湖南早晚稻生产和消费需求提出的育种设想,即在湖南同一个稻区,同一个品种通过早稻、晚稻“双审定”,且两季总生育期不超过220.0 d、两季总产量超过1000.0 kg/667 m^(2),早季米质达部标优质3级及以上,晚季米质达部标优质2级及以上。据此育成的水稻新品种松雅7号具有株型紧凑、茎秆粗壮、耐密植、芽期耐低温低氧能力较强等特点,在湖南省作早、晚双季稻种植生育期适宜、米质较好、丰产稳产,适宜机插、直播等轻简栽培。2023年通过湖南省早稻审定,2024年通过湖南省晚稻审定。本文介绍了松雅7号的选育经过和特征特性,及“双季同种”的概念、品种选育思路和具体要求,讨论了“双季同种”生产模式的实际意义。

水稻群体分蘖动态模型构建与应用

为定量分析水稻群体茎蘖数量动态变化过程及分蘖动态特征,该研究使用双Logistic模型分别描述分蘖发生与死亡过程,建立水稻群体分蘖动态模型;根据水稻分蘖过程的时序特征定义描述分蘖过程的特征指标,并推导出分蘖特征指标的计算式;基于不同基因型品种的种植方式、种植时期、种植密度下水稻分蘖动态数据集检验模型优度和适应性;并应用分蘖动态模型和指标探索分蘖动态对种植密度的响应规律。结果表明,所建模型对不同基因型水稻品种在不同种植方式、种植时期和种植密度下的分蘖动态数据拟合优度较好,标准均方根误差S_(RMSE)服从均值小于5%的Gamma分布,并且99%的S_(RMSE)小于10%。基于所建模型计算的分蘖特征指标(包括模型参数)对种植密度有很好的响应;留一法检验表明模型的预测性较好,观测值与模拟值的R^(2)=0.96。所建模型能够精确描述水稻茎蘖数量演变过程,具有很好的拟合优度、适应性和可解释性,可用于分析基因、环境、农艺措施对分蘖动态的影响,分蘖特征指标可望成为分析基因与环境互作的重要表型参数,对指导水稻精准栽培也有重要理论价值和实际意义。

豫北两熟区不同夏播作物对后茬冬小麦中后期群体微环境及产量的影响

为探讨不同夏播作物前茬对后茬冬小麦生长和产量的影响,在河南省新乡市获嘉县开展田间二因素试验,设置两个不同施肥水平(常规施肥和不施氮肥)和三种夏播作物前茬(玉米、大豆和花生),分析小麦拔节期至成熟期叶面积指数(LAI)、叶夹角(LA)、光合有效辐射(PAR)、叶片叶绿素含量(SPAD)、叶绿素荧光(F_(v)/F_(m)),以及土壤含水量(SWC)、土壤温度(Ts)和土壤呼吸速率(Rs)变化,并在成熟期测定小麦产量及其构成因素。结果表明,与常规施肥(CK)相比,不施氮肥条件下不同前茬的小麦LA略增,而LAI、IPAR、SPAD、F_(v)/F_(m)、SWC、Ts和Rs均有所降低。不施氮肥条件下,花生前茬的小麦LAI、IPAR、SPAD、F_(v)/F_(m)、SWC、Ts和Rs均显著低于其他两种前茬(P<0.05),其中花生、大豆和玉米前茬的SWC较CK的降幅分别为13.56%、13.39%和10.77%。与CK相比,不施氮肥条件下,花生前茬的小麦LA较CK的增幅在3种前茬中居中,小麦株型较为紧凑,有利于改善植株下层光合辐射,为群体生长创造良好的光合和土壤条件,其产量较玉米和大豆前茬分别高20.0%和21.9%;玉米前茬的整体表现不如大豆和花生前茬,但其和花生前茬的小麦千粒重和群体数均较高,因而产量居中;大豆前茬的小麦结实小穗数和干物质量较高,但其小麦千粒重较低,最终籽粒产量也较低。综合来看,在两熟种植模式中,选用花生作为冬小麦的前茬作物是豫北地区一年两熟区作物配置的一种较优选择。

双壳体核电厂空间减震结构的竖向动力试验研究

为降低核电厂内安全壳的竖向加速度,提出了一种双壳体空间减震结构,基于该结构的力学特性建立了简化三质点三自由度动力模型,进一步给出了结构响应传递函数并进行了参数分析,明确了结构质量比、阻尼比和频率比对结构竖向地震响应的影响规律,完成了双壳体空间减震结构的缩尺振动台动力试验。结果表明:双壳体空间减震结构可减小内安全壳的竖向加速度,减震率为12.23%~27.84%。双壳体空间减震结构的振动台试验响应结果与理论计算值吻合较好,验证了所提出的双壳体空间减震结构简化模型的准确性以及双壳体空间减震系统的有效性。

高接低栽式差速变姿态行星轮系栽植机构设计与试验

针对目前旱地蔬菜钵苗移栽机大多适用于大株距(大于260 mm)移栽且投接苗与栽植位置作业高度差大、移栽性能不佳等问题,本文提出一种高接低栽式差速变姿态行星轮系栽植机构。根据中小株距蔬菜钵苗移栽农艺指导与机构设计要求,解析双行星架差速行星轮系栽植机构工作原理并建立其运动学理论模型;结合所提出优化目标构建目标函数,开发了基于Matlab GUI的栽植机构计算机辅助分析优化设计软件,通过人机交互方式获得1组较优的机构设计参数组合。通过三维建模与装配以及ADAMS软件虚拟仿真,初步验证了机构正确性与合理性。开展栽植机构物理样机试制与试验台架系统开发研究,通过空转试验测试,验证了栽植机构实际作业轨迹、姿态与虚拟仿真及理论设计的一致性;开展栽植机构接苗与栽植性能试验,试验结果表明,栽植机构在接苗与栽植各项试验指标均较为优秀,能够满足栽植机构预期设计要求与旱地移栽机械标准,验证了该机构的可行性与实用性。

安徽稻油两熟制不同种植方式下气候资源配置和演变

安徽沿江地区稻-油复种模式下茬口衔接紧密、光温资源紧张,季节间气候资源配置备受关注。移栽和直播是补充和调配气候资源的有效种植方式,而稻油两熟制中不同种植方式下不同作物生长季气候资源配置、演变特征以及对未来气候变化的适应性尚不清晰。本研究以安徽沿江地区27个气象站1992—2022年的气温、日照时数、总辐射和降水等气象资料为基础,分析了不同种植方式下稻-油复种季节间气候资源配置与演变特征以及光温生产潜力。结果表明,近30年稻-油复种模式下季节间总辐射量、日照时数和光合生产潜力均呈下降趋势,气温、降水量和光温生产潜力呈上升趋势。水稻季移栽和直播方式下总辐射量倾向率分别为-27.9 MJ·m^(-2)·(10a)^(-1)和-28.8MJ·m^(-2)·(10a)^(-1),油菜季分别为-40.5 MJ·m^(-2)·(10a)^(-1)和-26.6 MJ·m^(-2)·(10a)^(-1)。水稻季移栽和直播方式下平均日最高气温倾向率分别为0.30℃·(10a)^(-1)(P<0.05)和0.24℃·(10a)^(-1),最高达32.70℃(2022年,青阳)。油菜季移栽和直播方式下平均气温倾向率分别为0.36℃·(10a)^(-1)(P<0.01)和0.39℃·(10a)^(-1)(P<0.01),移栽方式下平均气温较直播方式高0.96~1.43℃。稻油两熟制两季均面临光资源持续下降和气温不断上升等问题,采用移栽方式可通过延长作物生育期优化光、温资源配置,提高光温生产潜力。同时,适当推迟移栽期将有利于应对油菜苗期和水稻花期的高温。直播方式下宜采用高光效品种和构建光资源高效利用作物群体。

水分调控下绿肥种植和石灰施用对双季稻稻米镉含量的影响

[目的]农田镉(Cd)污染是当前我国水稻产业高质量发展面临的主要环境问题之一,严重影响稻米商品价值和人民健康。本研究采用田间大区试验,探究水分调控下绿肥种植、石灰施用对稻米Cd含量及水稻产量的影响,以期为水稻安全生产提供技术参考。[方法]试验用早、晚稻品种分别为湘早籼24号和华润2号,试验地在湖南省益阳市,设置常规栽培(CK)、水分调控(W)、绿肥+水分调控(GW)、石灰+水分调控(LW)、绿肥+石灰+水分调控(GLW)共5个处理。[结果]与CK处理相比,W、GW和LW处理早晚稻平均糙米镉含量分别显著降低34.5%、83.5%和83.3%,稻谷镉含量分别显著降低26.3%、82.5%和83.4%;GLW处理仅晚稻的糙米镉含量显著降低43.7%,稻谷镉含量显著降低40.8%。相比CK处理,GW和LW处理早晚稻平均稻壳镉含量分别显著降低71.3%和74.2%,茎叶镉含量显著下降85.3%和79.1%;GLW处理仅晚稻的稻壳镉含量显著降低38.5%,茎叶镉含量显著降低51.9%。相比CK处理,W、GW和LW处理早晚稻平均灌浆期土壤pH值分别显著提高6.2%、23.7%和20.0%,GW处理早晚稻灌浆期土壤有效镉含量显著降低20.0%~22.6%。相比CK处理,GW和LW处理可显著降低早稻稻壳中的Cd向糙米转运;除W处理早稻稻壳和晚稻茎叶较CK处理差异不显著外,其余各处理均可显著降低土壤中的Cd向茎叶、糙米和稻壳富集。各处理间水稻产量差异不显著。[结论]在水稻孕穗后淹水灌溉基础上,配套冬季豆科绿肥种植或春季石灰施用,可实现轻度镉污染稻田水稻安全生产,不影响水稻丰产。

基于质构仪TPA模式构建蛋白肠感官评分预测模型及应用

为建立一套客观、数据化评价动植物双蛋白肠感官综合评分的方法,该研究采用质构剖面分析(TPA)测定24组不同配方的样品,结合感官评价方法,通过逐步回归分析建立感官综合评分预测模型,并将其应用于响应面实验以优化双蛋白肠的配方。结果表明,动植物双蛋白肠的感官综合评分预测模型为Y=-68.110+0.003×硬度+79.119×弹性-0.001×咀嚼性-18.393×回复性,该模型相关系数R~2为0.902,校正决定系数R_(Adj)~2为0.875;响应面回归模型相关系数R~2为0.962,校正决定系数R_(Adj)~2为0.924,预测系数R_(Pred)~2为0.861,优化后蛋白肠的最佳配方为大豆拉丝蛋白质量分数74%、猪肉质量分数16%、大豆分离蛋白与花生蛋白粉比例7∶3、猪肉肥瘦比例7∶3,该配方下产品的感官评分实际值为8.51,与理论预测值(8.45)误差较小。该研究得到的动植物双蛋白肠感官综合评分预测模型的拟合度较高,实测值与预测值的吻合度较好,模型在响应面实验中具有可行性,为客观、快捷地评价新型肉肠的感官品质提供了新思路。

双链RNA对薇甘菊根基因 MmEXPA4 的靶向抑制

【目的】结合薇甘菊较强的节节生根能力的生物学特性,研发利用RNAi技术抑制薇甘菊根系生长基因表达的生物防治技术,为进一步开发薇甘菊靶向防控技术奠定基础。【方法】利用同源比对方法,鉴定出调控薇甘菊根系发育的关键基因(MmEXPA4),体外合成该基因的双链RNA(dsMmEXPA4),利用dsMmEXPA4注射薇甘菊根部,研究其对薇甘菊根系生长的作用。【结果】与对照组相比,dsMmEXPA4处理30 d后,所有不定根的数量、鲜重和最长不定根的长度均显著降低。qRT-PCR检测结果表明,在dsMmEXPA4处理后第4和第5天,MmEXPA4基因的表达量显著下调,沉默效率分别为43.96%和52.11%。此外,通过Fluorescent Stereo Microscope Leica M165 FC发现,被荧光标记的dsMmEXPA4能在根系中测检到较强的绿色荧光信号。【结论】MmEXPA4基因可作为抑制薇甘菊根系发育及快速生长的潜在靶点,为利用RNA干扰技术生物防治薇甘菊提供理论基础。

“绿茶+红茶”双茶联动加工模式研究--以瑞安市岭雅绿茶综合开发有限公司为例

求与精细工艺流程,以及红茶加工的采摘标准与小种红茶制作工艺。最后,介绍该基地采用的新型绿色防控技术与有机肥料应用情况。双茶联动加工模式提高资源利用效率,为茶叶生产实现可持续发展提供经验借鉴。

“双碳”目标下潘集选煤厂发展方向探索及实践

介绍了“双碳”目标下选煤厂的发展方向,绿色、高效、节能、智能化是选煤厂发展的主旋律。潘集选煤厂积极响应“双碳”目标号召,结合生产实际,从降低电耗、减少煤泥量、加强智能化建设三个方面进行工艺优化改进,取得了良好的经济和社会效益。

计算机数据融合算法下植保机精准化作业研究

针对我国传统的植保机在精准化作业和自动化喷洒方面存在的不足,基于计算机数据融合算法对植保机精准化作业进行了设计试验。植保机的主要组成包括控制单元模块、信息采集模块、传感器模块、数据传输模块和变量执行器模块。为了实现植保机的精准化作业,对植保机的飞行姿态调整和变量喷药过程进行了算法设计。其中,飞行姿态控制采用双闭环位置式PID控制算法,喷药过程采用改进的遗传PID控制算法进行控制。为了验证植保机精准化作业的效果,对其进行姿态控制和变量喷药试验。结果表明:植保机姿态控制良好,能够完成精准化喷药。