摘要：【中文】本发明公开一种基于烟碱定量转化率和基因表达筛选朱砂烟种子的方法，包括从朱砂烟种子库中选种育苗；出苗到生长30～40d且长至30～40cm高后选底部第2至第4片状态良好烟叶取样；将取样置于乙烯利+脱落酸混合溶液中浸润后打孔，将打孔后剩余烟叶干燥至表面无水分；将无水分烟叶烘干至恒重；用化学分析烟叶的烟碱和降烟碱含量，计算烟碱转化率；检测打孔烟叶CYP82E4基因表达；根据烟碱转化率和CYP82E4基因表达，筛选出符合烟碱定量转化率的烟株培植，同时去掉转化率不符烟株；将筛选出的烟株套袋至成熟留种，即得到烟碱定量转化率的朱砂烟种子。本发明具有稳定可靠、筛选速度快、准确率高、种子烟碱转化率可控等特点。 【EN】The invention discloses a method for screening cinnabar tobacco seeds based on nicotine quantitative conversion rate and gene expression, which comprises the steps of selecting seeds from a cinnabar tobacco seed bank and growing seedlings; selecting good tobacco leaves in the sheet states from the No. 2 to the No. 4 at the bottom for sampling after the seedlings emerge and grow for 30-40 days and grow to the height of 30-40 cm; placing the sample in a ethephon + abscisic acid mixed solution for soaking, then punching, and drying the residual tobacco leaves after punching until the surface is free of moisture; drying the moisture-free tobacco leaves to constant weight; chemically analyzing the nicotine and nicotine reducing content of the tobacco leaves, and calculating the nicotine conversion rate; detecting punched tobacco leavesCYP82E4Gene expression; according to the nicotine conversion rate andCYP82E4gene expression, screening out tobacco plants which accord with the nicotine quantitative conversion rate for cultivation, and removing tobacco plants with the conversion rate which does not accord with the nicotine quantitative conversion rate; bagging the screened tobacco plants until the tobacco plants are mature and reserved, and obtaining the nicotine quantitative conversion rateCinnabaris seed of Nicotiana cinnabarina. The method has the characteristics of stability, reliability, high screening speed, high accuracy, controllable seed nicotine conversion rate and the like.

摘要：【中文】本发明公开了一种预测烟草果糖含量的全基因组选择模型及其应用，所述的用于预测烟草果糖含量的全基因组选择模型为Bayes C_FRC，为使该模型对烟草中果糖含量表型值的预测精度达到最优，对候选预测模型Bayes C中的分子标记数量（n1）、训练群体规模（n2）、训练群体与测试群体比例（n3）及模型预测精度值（n4）等核心参数值进行了明确规定。所述的应用为利用所述的全基因组选择模型Bayes C_FRC分析烟草群体的基因型数据来预测其果糖含量的应用。本发明所述的烟草果糖含量全基因组选择模型Bayes C_FRC可以依据烟草群体的基因型来精确的预测出该群体中各烟株的果糖含量值，从而实现烟草品质育种中不同果糖含量水平优良品种（系）的培育。 【EN】The invention discloses a whole genome selection model for predicting the fructose content of tobacco and application thereof, wherein the whole genome selection model for predicting the fructose content of tobacco is Bayes C_FRCIn order to optimize the prediction accuracy of the model on the phenotype value of the fructose content in tobacco, core parameter values such as the number of molecular markers (n 1), the scale of a training population (n 2), the ratio of the training population to a test population (n 3), the model prediction accuracy value (n 4) and the like in a candidate prediction model Bayes C are clearly specified. The application is to use the whole genome to select a model Bayes C_FRCUse of genotypic data from a population of tobacco to predict its fructose content. The invention discloses a tobacco fructose content whole genome selection model Bayes C_FRCThe fructose content value of each tobacco plant in the tobacco population can be accurately predicted according to the genotype of the tobacco population, so that different fructose values in tobacco quality breeding are realizedCultivating the excellent variety (line) with the content level.

摘要：【中文】本发明公开了一种预测烟草淀粉含量的全基因组选择模型及其应用，所述的用于预测烟草淀粉含量的全基因组选择模型为rrBLUP_st，为使该模型对烟草中淀粉含量表型值的预测精度达到最优，对经过初步筛选而获得的候选预测模型rrBLUP中的分子标记数量（n1）、训练群体规模（n2）、训练群体与测试群体比例（n3）及模型预测精度值（n4）等核心参数值进行了明确规定。所述的应用为利用所述的全基因组选择模型rrBLUP_st分析烟草群体的基因型数据来预测其淀粉含量的应用。本发明所述的烟草淀粉含量全基因组选择模型rrBLUP_st可以依据烟草群体的基因型来精确预测出该群体中各烟株的淀粉含量值，从而实现烟草品质育种中不同淀粉含量优良烟草品种（系）的培育。 【EN】The invention discloses a whole genome selection model for predicting the content of starch in tobacco and application thereof, wherein the whole genome selection model for predicting the content of starch in tobacco is rrBLUP _ st, and in order to ensure that the model can achieve the optimal prediction precision on the phenotypic value of the content of starch in tobacco, core parameter values such as the number of molecular markers (n 1), the scale of a training population (n 2), the proportion of the training population to a testing population (n 3) and the model prediction precision value (n 4) in a candidate prediction model rrBLUP obtained through preliminary screening are definitely specified. The application is the application of utilizing the whole genome selection model rrBLUP _ st to analyze the genotype data of a tobacco population to predict the starch content of the tobacco population. The tobacco starch content whole genome selection model rrBLUP _ st can accurately predict the starch content value of each tobacco plant in a tobacco population according to the genotype of the population, thereby realizing the cultivation of tobacco varieties (lines) with excellent starch content in tobacco quality breeding.

摘要：【中文】本发明公开了一种水生产碱菌及其获取方法与应用，所述的水生产碱菌为水生产碱菌（Alcaligenes aquatilis）05‑101，已在中国典型培养物保藏中心保藏，其保藏编号为CCTCC No.M2019665。获取方法包括分离、筛选程序，首先从烟株组织中分离能降解尼古丁的菌株，再用以NNK为唯一碳源和氮源的培养基筛选即得。菌株的应用包括发酵、接种、降解工序，首先将菌株在28~30℃下培养36~72 h得发酵菌剂，于烟叶调制或制丝过程中，将发酵菌剂按烟草重量的3~5%喷施，保持4~7 d（烤烟）、45~50 d（晾烟）的降解时间即可。所述菌株能实现烟草特有亚硝胺的高效定向降解，且抗逆性好，简便易得，使用方便，具有较高的推广应用价值。 【EN】The invention discloses an Alcaligenes aquaticus as well as an acquisition method and application thereof, wherein the Alcaligenes aquaticus is Alcaligenes aquatilis 05-101 which is preserved in China center for type culture preservation with the preservation number of CCTCC No. M2019665. The acquisition method comprises the steps of separation and screening, wherein bacterial strains capable of degrading nicotine are separated from tobacco plant tissues, and then the bacterial strains are screened by using NNK as a unique carbon source and a unique nitrogen source. The application of the strain comprises the working procedures of fermentation, inoculation and degradation, the strain is cultured for 36-72 hours at 28-30 ℃ to obtain a fermentation inoculum, the fermentation inoculum is sprayed according to 3-5% of the weight of tobacco in the tobacco leaf modulation or shredding process, and the degradation time of 4-7 d (flue-cured tobacco) and 45-50 d (air-cured tobacco) is kept. The strain can realize the efficient directional degradation of the nitrosamine specific to the tobacco, has good stress resistance, is simple, convenient and easy to obtain, is convenient to use, and has higher popularization and application values.

摘要：【中文】本发明公开了一种高地芽孢杆菌J45及其获取方法与应用，所述的高地芽孢杆菌为高地芽胞杆菌（Bacillus altitudinis）J45，已在中国典型培养物保藏中心保藏，其保藏编号为CCTCC No.M2019666。获取方法包括分离、筛选、纯化工序，首先从烟株组织中分离能降解尼古丁的菌株，再用以尼古丁为唯一碳源的NIM培养基筛选，最后在培养基平板上纯化即得。菌株的应用包括发酵、接种、降解工序，首先将菌株在30℃下培养48~72h得发酵菌剂，于烟叶调制或制丝过程中，将发酵菌剂按烟草重量的3~5%喷施，保持4~7天的降解时间即可。所述菌株能实现烟草特有亚硝胺的高效定向降解，且抗逆性好，简便易得，使用方便，具有较高的推广应用价值。 【EN】The invention discloses a bacillus altitudinis J45 and an acquisition method and application thereof, wherein the bacillus altitudinis is bacillus altitudinis (Bacillus altitudinis)Bacillus altitudinis) J45, which has been preserved in China center for type culture Collection with the preservation number of CCTCC No. M2019666. The method comprises the steps of separating, screening and purifying, wherein bacterial strains capable of degrading nicotine are separated from tobacco plant tissues, then NIM culture medium with nicotine as a unique carbon source is used for screening, and finally, the bacterial strains are purified on a culture medium flat plate. The application of the strain comprises the working procedures of fermentation, inoculation and degradation, firstly, the strain is cultured for 48-72 hours at 30 ℃ to obtain a zymogen agent, the zymogen agent is sprayed according to 3-5 percent of the weight of tobacco in the process of tobacco leaf modulation or shredding,and keeping the degradation time for 4 to 7 days. The strain can realize the efficient directional degradation of the nitrosamine specific to the tobacco, has good stress resistance, is simple, convenient and easy to obtain, is convenient to use, and has higher popularization and application values.

摘要：【中文】本发明公开了烟草多酚氧化酶基因NtPPO2a和/或NtPPO2b在抗旱烟草品种选育中的应用，NtPPO2a核苷酸序列如SEQ ID：No.1所示，NtPPO2b核苷酸序列如SEQ ID：No.2所示。NtPPO2a编码的氨基酸序列如SEQ ID NO：3所示；NtPPO2b编码的氨基酸序列如SEQ ID：No.4所示。烟草多酚氧化酶基因NtPPO2a和/或NtPPO2b的定点突变方法包括载体构建与pORE‑CRISPR/Cas9表达载体的农杆菌转化及烟草的遗传转化等步骤。利用实时荧光定量PCR对烟草品种进行检测分析，当烟草多酚氧化酶基因NtPPO2a和/或NtPPO2b发生突变时，该品种抗旱性能降低。本发明为抗旱烟草品种选育提供了基础材料与检测方法。 【EN】The invention discloses a tobacco polyphenol oxidase geneNtPPO2aAnd/orNtPPO2bThe application in the breeding of drought-resistant tobacco varieties,NtPPO2athe nucleotide sequence is shown as SEQ ID: as shown in the No.1, the main component,NtPPO2bthe nucleotide sequence is shown as SEQ ID: no. 2.NtPPO2aThe coded amino acid sequence is shown as SEQ ID NO: 3 is shown in the specification;NtPPO2bthe coded amino acid sequence is shown as SEQ ID: no. 4. Tobacco polyphenol oxidase geneNtPPO2aAnd/orNtPPO2bThe site-directed mutagenesis method comprises the steps of constructing a vector, carrying out agrobacterium transformation on the pORE-CRISPR/Cas9 expression vector, carrying out genetic transformation on tobacco and the like. Detecting and analyzing tobacco varieties by using real-time fluorescent quantitative PCR (polymerase chain reaction), and detecting and analyzing tobaccoPolyphenol oxidase geneNtPPO2aAnd/orNtPPO2bWhen mutation occurs, the drought resistance of the variety is reduced. The invention provides a basic material and a detection method for breeding drought-resistant tobacco varieties.

摘要：【中文】本发明公开了一种温度振荡的烟叶调制工艺方法，属于烟叶调制技术领域。工艺方法依次包括变黄期、凋萎期、定色期及干筋期；变黄期包括起始升温速度振荡和变黄期温度振荡工序；凋萎期包括凋萎期升温速度振荡和凋萎期温度振荡工序；定色期定色期升温速度振荡和定色期温度振荡工序；干筋期为后续工序，干球温度以1℃/h的速度，将干球温度升至67.0～68.0℃，湿球温度为40.0～42.0℃，直至全炉烟叶干燥停止；升温速度振荡工序具体为在升温期间的每小时升温速度遵循不等同振荡的工序；温度振荡工序在稳温期间遵循升温、降温交替进行的工序。采用本发明所述工艺方法，烤后烟叶的品质特征、风格特征及卷烟工业可用性，明显提升，并具有较强的稳定性。 【EN】The invention discloses a tobacco leaf modulating process method with temperature oscillation, and belongs to the technical field of tobacco leaf modulation. The process method sequentially comprises a yellowing stage, a withering stage, a color fixing stage and a tendon drying stage; the yellowing stage comprises the working procedures of initial temperature rise speed oscillation and yellowing stage temperature oscillation; the wilting period comprises the working procedures of temperature rise speed oscillation in the wilting period and temperature oscillation in the wilting period; a temperature rising speed oscillation and color fixing period temperature oscillation process in the color fixing period and the color fixing period; the stem drying period is a subsequent process, the dry bulb temperature is increased to 67.0-68.0 ℃ at the speed of 1 ℃/h, the wet bulb temperature is 40.0-42.0 ℃ until the tobacco leaves in the whole furnace are dried; the temperature-rise rate oscillation step is a step in which the temperature rise rate follows unequal oscillation per hour during the temperature rise period; the temperature oscillation process follows a process of alternately increasing and decreasing the temperature during the temperature stabilization period. By adopting the process method, the quality characteristics and style characteristics of the flue-cured tobacco leaves and the industrial availability of cigarettes are obviously improved, and the process method has stronger stability.

摘要：【中文】本发明公开了一种烤烟滴灌水肥一体化方法，包括以下步骤：1）铺设滴灌带：烟株移栽前单垄单行铺设滴灌带，滴头距离烟株生长位置9‑11cm；2）计算水溶肥用量：根据土壤肥力状况确定待施入水溶肥的总用量；3）根据烟苗生长期分7次滴灌不同浓度不同用量的水溶肥；本发明追肥阶段均采用水肥一体化的滴灌施肥，有效提高了肥料利用效率，同时控制化学肥料投入，增加烤烟生产效益，提升了烟叶品质及其工业可用性；显著减少了施肥过程中的人工投入，大大提高了施肥效率。 【EN】The invention discloses a drip irrigation water and fertilizer integration method for flue-cured tobacco, which comprises the following steps: 1) laying a drip irrigation tape: before the tobacco plants are transplanted, single-ridge single-row drip irrigation belts are laid, and the distance between a dripper and the growth position of the tobacco plants is 9-11 cm; 2) calculating the dosage of the water-soluble fertilizer: determining the total amount of the water-soluble fertilizer to be applied according to the soil fertility condition; 3) drip irrigation water soluble fertilizers with different concentrations and different dosages for 7 times according to the growing period of tobacco seedlings; according to the invention, water and fertilizer integrated drip irrigation fertilization is adopted in the topdressing stage, so that the utilization efficiency of the fertilizer is effectively improved, meanwhile, the input of chemical fertilizer is controlled, the production benefit of flue-cured tobacco is increased, and the quality and industrial availability of tobacco leaves are improved; the labor input in the fertilization process is obviously reduced, and the fertilization efficiency is greatly improved.

摘要：【中文】本发明公开了一种烤烟滴灌水肥一体化肥料，分为基肥和追肥，基肥包括复混肥（N‑P₂O₅‑K₂O=10‑12‑24）和普钙（过磷酸钙，P₂O₅≥16%），追肥包括硫酸钾（K₂O≥50%）和硝酸铵钙（N≥15%）；本发明还公开了一种烤烟滴灌水肥一体化肥料的施肥方法，采用先施基肥后施追肥的方法，控制氮肥基肥追肥比为3:7～5:5，追肥分两次施用，第一次追肥的量为总追肥量的30~50%。本发明优化了施肥的氮磷钾比例，在一定程度上减少了肥料用量，降低了氮肥成本投入；本发明基于较低的施肥成本和较少的肥料用量，即实现了优质烟叶原料生产，从而提高了烟叶的经济效益；本发明追肥阶段均采用水肥一体化的滴灌施肥，显著减少了施肥过程中的人工投入，进一步提高了施肥效率。 【EN】The invention discloses a drip irrigation water-fertilizer integrated fertilizer for flue-cured tobacco, which comprises a base fertilizer and an additional fertilizer, wherein the base fertilizer comprises a compound fertilizer (N-P)₂O₅‑K₂O = 10-12-24) and superphosphate (calcium superphosphate, P)₂O₅Not less than 16%), and additional fertilizer including potassium sulfate (K)₂O is more than or equal to 50 percent) and calcium ammonium nitrate (N is more than or equal to 15 percent); the invention also discloses a method for applying the water-fertilizer integrated fertilizer for flue-cured tobacco by drip irrigation, which is characterized in that a method of applying base fertilizer first and then applying additional fertilizer is adopted, the ratio of the base fertilizer to the additional fertilizer of the nitrogen fertilizer is controlled to be 3:7-5:5, the additional fertilizer is applied for two times, and the amount of the first additional fertilizer is 30-50% of the total additional fertilizer. The invention optimizes the proportion of nitrogen, phosphorus and potassium for fertilization, reduces the fertilizer dosage to a certain extent, and reduces the cost input of the nitrogen fertilizer; the method realizes the production of high-quality tobacco raw materials based on lower fertilization cost and less fertilizer consumption, thereby improving the economic benefit of the tobacco; the invention adopts water and fertilizer integrated dripping in the topdressing stageAnd the fertilizer is applied by irrigation, so that the labor input in the fertilizing process is obviously reduced, and the fertilizing efficiency is further improved.

摘要：【中文】本发明公开了一种基于窄株宽行栽培的烤烟高效灌溉方法，采用株间交替灌溉的方式进行灌溉，设定每行种植n株烟苗，n为大于或等于7的整数，整个生育期具有m个灌溉期，m为大于等于3的整数；第m次灌溉：当m为单数时，灌溉第1株和第2株之间中心位置、第3株和第4株之间中心位置、第5株和第6株之间中心位置......第n‑3株和第n‑2株之间的中心位置，第n‑1和第n株之间的中心位置；当m为双数时，灌溉第2株和第3株之间中心位置、第4株和第5株之间中心位置、第6株和第7株之间中心位置......第n‑4株和第n‑3株之间中心位置、第n‑2和第n‑1株之间的中心位置。本发明在保障烟株生长发育的前提下，减少了烟田灌溉水用量，提高了烤烟灌溉效益。 【EN】The invention discloses a flue-cured tobacco efficient irrigation method based on narrow-plant wide-row cultivation, which is characterized in that the method of alternating irrigation among plants is adopted for irrigation, n tobacco seedlings are planted in each row, n is an integer greater than or equal to 7, the whole growth period has m irrigation periods, and m is an integer greater than or equal to 3; and (3) irrigation for the m time: when m is singular, irrigating the central positions between the 1 st and 2 nd plants, between the 3 rd and 4 th plants, and between the 5 th and 6 th plants, the central positions between the n-3 rd and n-2 nd plants, and the central positions between the n-1 th and n-2 th plants; when m is a double number, the center position between the 2 nd and 3 rd plants, the center position between the 4 th and 5 th plants, and the center position between the 6 th and 7 th plants are irrigated. The invention reduces the amount of irrigation water used in tobacco fields and improves the irrigation benefit of flue-cured tobacco on the premise of ensuring the growth and development of tobacco plants.