Detection and Quantitative Analysis of Dominant Bacteria On Aging Flue-cured Tobacco Leaves

[Objective] The aim was to construct the internal motivation of flued-cured tobacco natural aging. [Method] The variation of DNA copies of Bacillus megaterium, amylase activity, PPO, peroxydase and content of starch, neutro-protein, polyphenol in different aging period were investigated using Henan flue-cured tobacco leaves. [Result] Results showed that the DNA copies of Bacillus megaterium increased initially but decreased subsequently with the aging time. The DNA copies of Bacillus per gram of tobacco leaf surface reached the maximum number at 6 months aging. The activities of four enzymes showed the same variation trend as in DNA copies, and also reached the highest level at 6 months aging. Chemical ingredients, however, displayed a downward trend. The variation of enzyme activity and content of chemical composition were reflected in the changes of DNA copies in Bacillus megaterium. [Conclusion] The result indicate that Bacillus megaterium was the internal motivation of flue-cured tobacco natural aging.

Effects of Different Transplanting and Harvest Dates on Yield and Quality of Flue-cured Tobacco Leaves

In this study,the effects of nine different combinations of transplanting and harvest dates on yield and quality of flue-cured tobacco leaves were investigated.The results showed that different combinations of transplanting and harvest dates had few effects on growth period and agronomic characters of tobacco plants but exerted remarkable effects on economic characters and chemical composition of flue-cured tobacco leaves.With the postponing of transplanting stage,output value of flue-cured tobacco leaves increased.Output value of flue-cured tobacco leaves in C1treatment（normal harvest） was higher than that in C2（harvest 7 d later） and C3（harvest 14 d later） treatments.Different combinations of transplanting and harvest dates had significant effects on chemical composition of flue-cured tobacco leaves,which exhibited no obvious law.Overall,flue-cured tobacco leaves in T3C1treatment（transplanting 14 d earlier,normal harvest） displayed excellent performance compared with other treatments.

Comparative Analysis on Chemical Components and Sensory Quality of Aging Flue-Cured Tobacco from Four Main Tobacco Areas of China

Complicated changes are happening during the aging process of flue-cured tobacco （FCT） and understanding of the changing components is of great significance in improving the quality,shortening aging time,and enhancing production and economic efficiency in tobacco industry.The aging samples of FCT from four main producing areas of China,i.e.,Yunnan,Sichuan,Hubei,and Dongbei,were used to study the changing rule of the components such as alkali,acid,and carbohydrate as well as the aroma and their relationship with sensory quality;and based on the correlation among the components and the sensory quality index,multivariable models were established to predict the aging quality.The results showed that the sensory quality of FCT increased steadily during the aging time,and at the same time,the acidic components （total organic acids,volatile acids） increased gradually,while the alkaline substances （nicotine,volatile alkali）,carbohydrate （total sugars,reducing sugar） and pH values showed a downward trend.Correlation analysis found that the sensory quality and pH values were negatively correlated （P0.05）,while the sensory quality with total organic acids and aroma were positively correlated.The optimal model for predicting the quality of FCT was y=56.75-0.63x12＋50.09x2-13.27x22,（y：sensory quality;x1：pH;x2：total organic acids）.The average predicating error was 1.04% with maximum predicating error of 2.89% and the accuracy rate of above 97%.

Judgment of Aroma Types of Middle and Upper Flue-cured Tobacco Leaves Based on Stepwise Discriminant Analysis

[Objective] The aim of this study was to establish mathematical models for judging the aroma types of middle and upper flue-cured tobacco leaves according to the contents and proportions of aroma compositions. [Method] The aroma types of tobacco leaves were judged based on stepwise discriminant analysis, using 63 C3F and 65 B2F tobacco leaf samples from 13 tobacco producing regions in 11 provinces of China （Huili in Sichuan, Baokang in Hubei, Wulong in Chongqing, Lu- oyang in Henan, Zhucheng in Shandong, Wuyi Mountain in Fujian, Malong in Yun- nan, Chuxiong in Yunnan, Bijie in Guizhou, Liuyang in Hunan, Suiyang in Guizhou, Kaiyuan in Liaoning, Nanxiong in Guangdong） as calibration samples, and 67 aroma components as indices. And the Fisher discriminant functions were verified using 21 C3F and 19 B2F tobacco leaf samples. [Result] Variation coefficients of the propor- tions were lower than that of contents of most aroma components in middle and upper leaves of the samples, indicating that the proportions were more stable than contents of aroma components. The proportions of benzyl alcohol, solanone, β-dam- ascone, neophytadiene, farnesylacetone A, palmitic acid, thunbergol, methyl linole- nate and cembratriene-diol were all over 1% in both middle and upper leaves, al- though the dominant aroma components of the same aroma type varied between middle and upper leaves. Moreover, 11, 18, 7 and 11 aroma components were re- spectively introduced into the Fisher discriminant functions established based on the contents and proportions of middle and upper flue-cured tobacco leaves, which ex- hibited accuracy rates of 91.7%, 100%, 91.7% and 91.7% in the judgments of other tobacco leaf samples. The results revealed that the components those determined aroma types in middle leaves were obviously more than in upper leaves. In middle leaves, the accuracy rates of aroma type judgment could be improved by using the proportions rather than the contents of aroma components as indices. However, the functions based on the proportions and the contents of aroma components in upper leaves gave close accuracy rates. [Conclusion] The results of the study will provide references for identifying aroma types of flue-cured tobacco leaves in future work.

Relationship Between Hyperspectral Parameters and Physiological and Biochemical Indexes of Flue-Cured Tobacco Leaves

The experiment was set up for examining the physiological and biological indexes quickly and exactly, for obtaining information of tobacco-field fertilizing and tobacco growing. The ASD Field spec FR 2500 was used to measure spectra reflectance of flue-cured tobacco and the relationship between hyperspectral parameters and biochemical contents （total nitrogen, chlorophyll, carotenoid）, and physiological indexes （fresh weight, dry weight, moisture content） of flue-cured tobacco leaves was studied by correlation and stepwise regression statistic methods at different nitrogen and potassium levels. The results indicated that the spectra curves of different treatments had obvious rules and great diversities. There were high correlations between different types of spectra parameters and ten physiological and biochemical indexes of flue-cured tobacco leaves. Hyperspectral characteristic variables of ten physiological and biochemical indexes were found through stepwise regression, and SDr/SDb was the characteristic variable closest to seven biochemical contents. Simultaneously, the R2 and regression coefficient of equations reached 0.05 significant level and the equations had good estimating effects through the examination of other samples. Accordingly, this study suggested that the ten physiological and biochemical indexes could be estimated quickly by the estimating models, at the same time nitrogen-potassium fertilization and growth condition of flue-cured tobacco could be inspected.

Judgment of Aroma Types of Middle and Upper Flue-cured Tobacco Leaves Based on Proportions of Aroma Components

[Objective] This study aimed to establish mathematical models for judging the aroma types of flue-cured tobacco leaves from the upper and middle parts of plants. [Method] A total of 128 samples （63 C3F and 65 B2F） from 11 main tobac- co production provinces of China were selected as materials. Stepwise discriminant analysis was applied to samples with different aroma types and discriminant function was expressed with the proportions of 67 aroma components in total aroma con- stituents as the index. [Result] The ratio of most aroma components in clear and full aroma tobacco leaves was higher than that in middle aroma leaves. The ratios of 51, 43 and 40 aroma components of clear, middle and full aroma tobaccos were higher in upper leaves than that in middle leaves. Aroma components dominated certain aroma types differed between middle and upper leaves. The proportions of 18 and 11 aroma components in upper and middle leaves were led in the stepwise discriminant function respectively. Self-validation and cross-validation methods were applied to evaluate the original samples, and the accuracy rates reached 100% and 98.6% on middle leaves, 96.37% and 94.4% on upper leaves. The accuracy rates on some other samples reached 100% on middle leaves and 91.7% on upper leaves predicted with the model. [Conclusion] The ratio of aroma components as discriminant index could improve discriminant accuracy significantly in the middle and upper leaves. It could be used to analyze aroma types objectively, accurately and quickly.

Analysis of Growth Laws of Leaves of Flue-cured Tobacco in Various Periods

The growth laws of leaves of flue-cured tobacco in different positions in various periods were summarized. The results showed that the growth speed of new leaves of flue-cured tobacco increased constantly until their growth was the fastest, and then it reduced irreversibly. The length and width of leaves in each position increased fastest when they grew depending on photosynthates instead of or- ganic compounds from functional leaves. Therefore, it is concluded that the increases of leaf length, width and area, and dry matter, and the peak of photosynthetic rate appear successively during the process of leaves of flue-cured tobacco, and the leaf area correction coefficient of leaves of flue-cured tobacco is related to the shape of leaves, which are affected by leaf position and the variety of flue-cured tobacco. The study can provide scientific reference for further research on the growth laws of leaves of flue-cured tobacco.

Effects of Different Flue-curing Technologies on Microscopic Structure of Flue-cured Tobacco Leaves

[Objective] This study aimed to compare the effects of Chinese quality-improving fragrunce-increasing tobacco flue-curing technology, Chinese three- stage tobaeeo flue-curing technology and Zimbabwean tobaceo flue-euring technology on microscopie structure of flue-cured tobacco leaves to provide theoretical basis for tobacco flue-curing. [ Method ] Middle leaves of tobacco cultivar K326 were collected in Pengshui County of Chongqing City for flue-curing experiment using three flue-curing technologies. Leaf samples were collected regularly in the flue-curing process, to investigate the microscopic structure of flue-cured tobaceo leaves. [ Result] During three flue-curing processes, leaves, palisade tissues and sponge tissues shrank gradually. Three flue-euring processes exhibited significant differ- ences in the peak of tissue shrinkage ： microscopic structure of flue-cured tobacco leaves in Chinese three-stage tobacco flue-curing process shrank earliest, follower by Chinese quality-improving fragrance-increasing tobacco flue-curing process; flue-eured tobacco leaves in Zimbabwean tobacco flue-curing process presented the latest shrinkage. At 84 h post-curing, leaf thickness in three flue-curing processes showed a descend order of Chinese three-stage tobacco flue-curing process 〉 Chinese quality-improving fragrance-increasing tobacco flue-curing process 〉 Zimbabwean tobacco flue-curing process. Upper and lower epidermal cells in Zimb- abwean tobacco flue-curing process ruptured earlier than other two flue-curing processes; eventually, the majority of cells ruptured and mixed with palisade tissues and sponge tissues. In Chinese quality-improving fragranee-increasing tobacco flue-curing process, only a small number of epidermal cells in dried leaves ruptured. Among three flue-curing technologies, Zimbabwean tobacco flue-curing technology exhibited the greatest damage to epidermal cells, followed by Chinese three-stage tobaceo flue-curing technology; Chinese quality-improving fragrance-increasing tobacco flue-curing technology had the minimum damage to epidermal cells. Mese- phyll cross-section exhibited significant morphological changes in stomata. To be specific, at 0 - 12 h post-curing, stomata of tobacco leaves closed with slightly prominent stamatal apparatuses on upper and lower epidermis; at 24 -72 h post-curing, stomata of tobaceo leaves changed gradually from opening to closure with significantly prominent stomatal apparatuses on upper and lower epidermis; at 84 h post-curing, stomata of tobacco leaves closed, and the majority of stomatal appa- ratuses were significantly prominent. In Chinese quality-improving fragrance-increasing tobacco flue-curing process, only a small number of epidermal ceils ruptured at 84 h pest-flue-curing; palisade tissues and sponge tissues shrank almost simultaneously. Significant gaps were observed between palisade tissues and between pal- isade tissues and sponge tissues. Chinese quality-improving fragrance-increasing tobacco flue-curing technology exhibited lower disorder level compared with other two flue-curing technologies. [ Conclusion] Chinese quality-improving fragrance-increasing tobacco flue-curlng technology was conducive to maintaining the micro- scopic structure integrity of flue-cured tobacco leaves and obtaining high-quality flue-cured tobacco leaves.

Effects of Different Flue-curing Technologies on Submicroscopic Structure of Flue-cured Tobacco Leaves

[ Objective] This study aimed to compare the effects of low-temperature low-humidity tobacco flue-curing technology, moderate-temperature moderate- humidity tobacco flue-curing technology and moderate-temperature high-humidity tobacco flue-curing technology on submicroscopic structure of flue-cured tobacco leaves to provide theoretical basis for tobacco flue-curing. [ Method] Middle leaves of tobacco cuhivar K326 were collected in Pengshui County of Chongqing City for flue-curing experiment using three flue-curing technologies. Leaf samples were collected regularly in the flue-curing process, to investigate the submicroscopic structure of flue-cured tobacco leaves. [ Result] Morphology changes and structural rupture of chloroplasts occurred at different time during three flue-curing proces- ses. In low-temperature low-humidity flue-curing process, morphology changes and structural rupture of chloroplasts occurred late but rapidly; in moderate-tempera- ture moderate-humidity flue-curing process and moderate-temperature high-humidity flue-curing process, morphology changes and structural rupture of chloroplasts occurred early but slowly. Among these three flue-curing processes, the speed of changes in mitochondria morphology, cristae number and membrane rupture pres- ented an upward order of low-temperature low-humidity flue-curing process 〈 moderate-temperature high-humidity flue-curlng process 〈 moderate-temperature moderate-humidity flue-curing process. In low-temperature low-humldity flue-curing process, starch granules in chloroplasts ruptured after flue-curing for 36 -48 h ; in moderate-temperature moderate-humidity flue-curing process, starch granules in chloroplasts ruptured after flue-curing for 24 - 36 h; in moderate-temperature high-humidity flue-curing process, starch granules in chloroplasts ruptured after flue-curing for 12 - 24 h. [ Conclusion ] Using low-temperature low-humidity flue- curing technology could prolong the maintenance duration of the submicroscopic structure integrity of flue-cured tobacco leaves, which was conducive to fully decom- posing and transforming substances in tobacco leaves and obtaining high-quality flue-cured tobacco leaves.

The Quality Difference of Tobacco Leaves Between Different Flue-curing Processes in Honghe Tobacco-growing Area

[Objectives] This study was conducted to compare the effects of different curing processes,in order to optimize the local supporting intensive fluecuring process.[Methods] A comparative experiment was carried out in Mile,Luxi and Jianshui on the four-layer tobacco-packed intensive curing process(K1)and the eight-point precise and intensive curing process(K2).[Results]The results showed that according to the appearance evaluation of the tobacco leaves,K1 was higher than K2 in respective scores of color,maturity,leaf structure and defects and the total score,while K1 equaled K2 in the scores of identity,oil content,chroma and length.The differences in respective scores of the two processes were not significant.According to the results of sensory evaluation,the two tobacco enterprises got different evaluation results of unblended cigarettes made of the middle leaves due to different raw material requirements of the enterprises by different processes.K1 was higher than K2 in the sensory evaluation of China Tobacco Yunnan Industrial Co.,Ltd.,while it was lower than K2 in that of China Tobacco Zhejiang Industrial Co.,Ltd.,and the difference in the total score between the two processes was significant.From the content of chemical components,in the conventional chemical component contents including total sugar,reducing sugar,chlorine and starch,K1 was higher than K2,while in the contents of potassium,total phytonine,total nitrogen and petroleum ether extract,K1 was lower than K2,but the differences were not significant.K1 was lower than K2 in the contents of neutral aroma components including ketones,alcohols,aldehydes,heterocyclics,neophytadiene and total content,while K1 was higher than K2 in the contents of esters,phenols,but the respective differences were not significant.Except the significant difference in the total score in the sensory evaluation of China Tobacco Zhejiang Industrial Co.,Ltd.,there were no significant differences in other items in the appearance evaluation,sensory evaluation and chemical composition of tobacco leaves between K1 and K2.[Conclusions] The supporting intensive curing process can be further optimized on the basis of this study.

Effects of Different Baking Processes on Carotenoid Content in Upper Leaves of Fluecured Tobacco

[Objective] We aimed to improve the quality of flue-cured tobacco with K326 upper leaves and explore the optimal tobacco baking process in Chongqing tobacco-producing region. [Method] Four different baking processes （endogenetic force moisture-removing flue-curing technique, exogenetic force moisture-removing flue-curing technique, three-stage flue-curing process, three-stage and six-step flue- curing process） were adopted in this research to investigate the effects of different baking processes on carotenoid content of same-maturity degree upper leaves of the flue-cured tobacco K326. [Result] The results showed at the yellowing and color fixing stages, the carotenoids were most fully decomposed in endogenetic force moisture-removing flue-curing technique. The decomposed carotenoid content was lowest in exogenetic force moisture-removing flue-curing technique, for which the carotenoid content was highest （50.53 mg/g）. [Conclusion] At the yellowing and color fixing stages, exogenetic force moisture-removing flue-curing technique was most conducive to the carotenoid accumulation in tobacco baking process.

Effects of Exogenous Amylases and Metal Ions on the Amylase Specific Activities and Starch Degradation of the Upper Leaves of ‘KRK_(26)' during Flue-curing

Objective] The aim of this study was to investigate the effects of exoge-nous amylases and Ca2＋, Mn2＋ and K＋ on the amylase specific activities and starch degradation of the upper leaves of ＇KRK26＇ planted in Yunnan Province during flue-curing. [Method] The amylase specific activities and starch degradation of the leaves were determined by using spectrophotometry. [Result] The 8 U/g exogenous α-amy-lase could improve the specific activity of the leaf α-amylase at yel owing and color-fixing stages, but could not at stem-drying stage, and similarly, the 80 U/g exoge-nous β-amylase could improved the specific activity of the leaf β-amylase at the yel owing stage and the early period of color-fixing stage. The leaf starch could be enhanced to degrade by the exogenous α- or β-amylases and the enhancing effect of the former was stronger than that of the later. 1.50 mg/ml Ca2＋ improved the specific activity of the leaf （α＋β）-amylase mainly due to its enhancing effect on the leaf α-amylase, and increased the starch degradation. 4 mmol/L Mn2＋ inhibited the leaf α-amylase from yel owing to the early period of color-fixing and the β- and （α＋β）-amylases from the yel owing to the later period of color-fixing, but enhanced the leafα-amylase from the later period of color-fixing to the later period of stem-drying and the β- and （α＋β）-amylases at the later period of stem-drying. Meanwhile, Mn2＋ ham-pered the starch degradation during yel owing, but promoted it from the early period of color-fixing to stem-drying. 1 mg/ml K＋ enhanced the leaf α-, β- and （α＋β）-amy-lases during the yel owing stage, but lowered them from the early period of color-fix-ing to the later period of stem-drying, and always inhibited the leaf starch degrada-tion. [Conclusion] The exogenous α-, β- amylases and Ca2＋ of suitable concentra-tions could be used to treat the tobacco leaves before flue-curing to improve the leaf starch degradation during the curing.

Analysis of the Appearance Quality of Tobacco Leaves in the Tobacco-growing Areas of Qujing City

In order to set up the digitization indices system of appearance quality of flue-cured tobacco leaves in Qujing. According to the relevant standards,the quantitative analysis on appearance quality of 39 cutter and upper tobacco leaf samples which were collected from eight fluecured tobacco production regions of Qujing City,Yunnan Province were carried out in 2011,respectively. The results showed that the difference in evaluation score of appearance quality of flue-cured tobacco in eight regions was not significant. The tobacco leaves had high maturity,high oil content,loose structure and good appearance quality. The scores of appearance quality of flue-cured tobacco in Shizong County and Malong County are higher than other regions. The two regions are suitable for tobacco plantation. The preliminary digitization indices provide reference and guidance to tobacco production and standards system of tobacco leaves base.

产纤维素酶工程菌株的构建及其在醇化烟叶中的应用

烟叶中过高的纤维素含量使烟叶组织容易破碎,影响加工过程中烟叶的可塑性,使烟叶杂气变重等。为了获得产纤维素酶的优良菌株,实现醇化烟叶纤维素的有效降解,利用同源重组法成功构建了10株纤维素降解的枯草芽孢杆菌(Bacillus subtilis)工程菌株。通过刚果红平板筛选、羧甲基纤维素钠酶活、滤纸酶活及滤纸崩解率等检测,共筛选出C36、CM、KF和GH54株产纤维素酶能力较强的重组菌株。将醇化烟叶作为底物进行纤维素降解,发现重组菌株CM的产纤维酶效率最高,其羧甲基纤维素钠酶活和滤纸酶活分别为39.55和23.52 U·mL-1。结果表明,构建的重组菌株能够利用醇化烟叶中的纤维素产生纤维素酶,可为工业生产中醇化烟叶纤维素降解提供理论支撑。

醇化烟叶关键微生物类群与风格特色的相关性

分别选取清香型(云南玉溪)、浓香型(湖南郴州)和中间香型(贵州遵义)醇化1年的C2F烟叶,测定烟叶的主要化学成分;利用相似度检验分析烟叶微生物群落差异物种,随机矩阵理论构建分子生态网络;运用Mantel分析烟叶主要化学成分与微生物群落的相关性,结果表明:清香型、中间香型和浓香型烟叶微生物群落结构存在显著差异,浓香型烟叶细菌及真菌群落ɑ多样性指数最高;烟叶显著富集的关键细菌类群为假单胞菌属(Pseudomonas)、肠杆菌属(Enterobacter)和鞘氨醇单胞菌属(Sphingomonas);关键真菌类群为链格孢属(Alternaria)、根霉菌属(Rhizopus)和桑帕约氏酵母菌属(Sampaiozyma);以浓香型烟叶微生物分子生态网络规模最大,结构复杂且相对稳定。总糖与还原糖含量显著影响烟叶微生物群落的多样性与结构,清香型及中间香型烟叶内关键微生物类群均与总糖、还原糖含量呈显著正相关,而浓香型烟叶关键微生物类群均与总糖、还原糖含量呈显著负相关。

微生物酶制剂对上部烟叶醇化品质的影响

为改善烟叶醇化品质,以低次红花大金元上部烟叶为试材,将BioApex微生物酶应用于烟叶醇化中,比较醇化烟叶的常规化学成分、挥发性香气成分和感官质量的变化。结果表明,添加微生物酶制剂会增加除氯以外的其他常规化学成分含量;同时降低了两糖差,使烟叶成分更加协调。各微生物酶制剂处理烟叶中还原糖、水溶性糖含量与CK相比均显著增加,但醇化时间延长会显著降低烟碱和水溶性糖含量。微生物酶制剂处理增加了烟叶中芳香族氨基酸降解产物、类胡萝卜素降解产物、美拉德反应产物含量。与CK相比,0.5%、1.0%和2.0%微生物酶制剂处理(90 d)明显改善了烟叶的感官质量。综上所述,微生物酶制剂能够促进烟叶内含物质的转化,提升红花大金元上部烟叶的品质。该研究可为烟叶微生物酶制剂发酵工艺在工业中的应用提供理论基础。

贮存方式对云南产烟叶醇化的影响研究

以云南宣威BFE、XFE和云南腾冲BFE烟叶为研究对象,对不同贮存库(机械库、自然库、保湿库)和不同醇化时间(3个月、9个月、12个月、15个月、18个月)下3种云南产烟叶的常规化学成分和挥发性香味成分进行对比分析,并对挥发性香味成分进行主成分分析。结果表明:1)烟叶中还原糖和总糖含量整体呈下降趋势,分别下降了9%~19%和12%~22%;自然库中醇化烟叶的还原糖和总糖含量下降速度较慢,机械库和保湿库中醇化烟叶的还原糖和总糖含量下降速度较快,氯离子和烟碱含量基本无变化,糖碱比呈波动性变化,云南腾冲BFE和云南宣威BFE烟叶中糖碱比的波动范围为7~11,而云南宣威XFE烟叶中糖碱比的波动范围为15~20。2)随着醇化的进行,贮存库中3种烟叶的挥发性香味成分含量呈快速上升趋势,其中酸类、酚类、酮类等物质增加较明显,且自然库中3种烟叶的挥发性香味成分含量最低。3)3种烟叶均提取两个主成分,其累计贡献率均达到100%,且贮存库中3种烟叶的挥发性香味成分的主成分都呈现出明显差异。因此,云南宣威BFE和XFE这2种烟叶更适合在机械库中进行醇化,而云南腾冲BFE烟叶更适合在保湿库中进行醇化。

打叶复烤强度及陈化时间对烟叶挥发性有机酸的影响

为探究打叶复烤强度及陈化时间对挥发性有机酸含量的影响,通过检测不同打叶复烤强度、不同陈化时间烟叶的挥发性有机酸含量,对比分析了打叶复烤强度及陈化时间对挥发性有机酸含量及组成的影响,以及打叶复烤强度对烟叶陈化程度的影响。结果表明:1)打叶复烤强度、陈化均会显著影响烟叶中有机性挥发酸的含量及组成,打叶复烤强度的影响小于陈化。2)打叶复烤强度会显著影响烟叶陈化的程度,随着打叶复烤强度增强,各种挥发性有机酸含量的平均增幅降低,但挥发性有机酸的组成变化加剧。控制打叶复烤强度对提升陈化后烟叶的品质和可用性均有重要意义。

陈化期间烟叶香气成份消长规律的研究

以烤烟（NicotianatabacumL.）为试材，研究了陈代期间烟叶香气物质和主要化学成份含量的变化。结果表明，随着陈化时间的延长．烟叶中大多数香气物质成分的含量持续上升，尤其是低分子量的成份上升明显；而还原精、烟碱、总氮等化学成份的含量则逐渐下降。在不同的陈化阶段．烟叶中香气物质和化学成份在12个月以前含量变化最为激烈，之后趋于缓慢。对陈化期间烟叶的烟气特性进行了鉴定，结果表明，陈化时间在18～24个月时烟叶的内在品质最好。

应用16S rDNA克隆文库技术分析陈化烟叶细菌多样性

为较全面地分析陈化烟叶的细菌多样性,利用16S rDNA基因克隆文库技术分析了产自于福建的3个等级处于不同陈化阶段的12个烟叶样品中的细菌组成。采集C/S挑、B22、X21三个等级的陈化烤烟烟叶,陈化时间分别为3、6、12、24个月。实验结果表明,烤烟烟叶在陈化过程中共存在25种细菌,γ-变形菌纲(Gamma-proteobacteria)细菌种类最丰富,含有19种细菌,α-变形菌纲(Alpha-proteobacteria)含有3种细菌,芽孢杆菌纲(Bacilli)含有2种细菌,β-变形菌纲(Beta-proteobacteria)含有1种细菌。在12个烟叶样品中,肠杆菌属(Enterobacter)在8个样品中的比例最高,假单胞菌属(Pseudomonas)在2个样品中比例最高,泛菌属(Pan-toea)和克洛诺菌属(Cronobacter spp.)分别在样品B6和C3中比例最高。在陈化过程中,烤烟叶面细菌组成没有明显的变化规律。