Effects of Bamboo Vinegar on Seed Germination and Seedling Growth of Different Flue-cured Tobacco Varieties

Lingjie XU,Shuangjian YANG,2,Min DONG,Yuanyuan SHI,Xiangge DU*

1.China Agricultural University,Beijing 100193,China;2.Weining Branch of Bijie Tobacco Company,Bijie 551700,China

Bamboo vinegar is a kind of dark brown liquid generated from the sealed pyrolysis and condensation of bamboo wood or its processed remainders (Fang Kaiet al.,2007),its major ingredient is water,and it also contains more than 200 ingredients,such as organic acid,phenols,ketone,alcohol,aldehydes,etc.(Zhang Wenbiaoet al.,2001).Bamboo vinegarhas bacteriostatic and bactericidal effects (Sulaimanet al.,2005),which can increase the effect of pesticides (Li Xiaoronget al.,2005),thus it can be used as plant growth regulator(Zhang Wenbiaoet al.,2004)and soil conditioner(Wei Qianget al.,2006),and it also can be used for keeping flowers fresh (Yang Hui,2005)etc.At the float breeding stage of flue-cured tobacco,the seedling of tobacco is easy to have the diseases ofdamping-off,RhizoctoniasolaniKühn,Colletotrichum micotianaeAverna,Thielaviopsis basicola(Brek.et Br.)Ferr.etc.(Zhou Zhichenget al.,2009),and the insects of mole cricket,cutworm,wireworm,grub,etc.,which seriously reduce the survival rate of tobacco seedlings.Tobacco growers often use chemical agents to prevent diseases and pests,which not only has poor effects,but also brings adverse effects on ecologicalenvironment.Therefore, to produce ecological,high-quality,characteristic and safe tobacco leaves,green(ecological)controlling technique is the basis for ensuring the safe poroduction of tobacco leaves based on the rule of“Good A-gricultural Practices”(GAP).So the study on the effects of bamboo vinegar on seed germination and seedling growth of different flue-cured tobacco varieties is of great significance in improving the quality of tobacco seedlings and reducing the dosages of chemical pesticides as well as protecting the environment.In recent years,as a kind of green medicament which can not only sterilize but also kill pests,bamboo vinegar has been widely used in agricultural production,moreover,it obtains favorable effect in a wide range of vegetables and fruit.Hu Chunshuiet al.found that the control effect of 40%dimethoate mixed with 3%1000 times bamboo vinegar on red spider of citrus orchard could reach 99.5%in 2000.Wei Qiang(2006)showed that spraying 100 times bamboo vinegar could effectively control the germination of spores withPseudoperonospora cubensis(Berk.et Curt.)Rostov.,its field control efficiency was comparable to 72%600 times Curzate.Adding 400 times bamboo vinegar into pepper Juanyeling could effectively improve the control efficiency of virus disease of pepper(Sun Jianhuaet al.,2006).Spraying bamboovinegarcanpromotethe growth vigour of tomato plants and improve VCcontent and sugar-acid ratio,moreover,the effect of 300 times bamboo vinegar was the best(Zhang Linet al.,2010).Spraying 300-1 500 times bamboo vinegar on the foliage could obviously improve VCcontent of tomato and decrease the density of peach aphids(Yao Yuanet al.,2012).At present,the reports on the effects of spraying bamboo vinegaratthe seedling stage on seed germination and seedling growth of different fluecured tobacco varieties have been fewer.Bamboo vinegar with different dilution times was used for culturing the seeds of different flue-cured tobacco varieties and sprayed on the leaves of seedlings,its effects on seed germination and seedling growth were studied,and the suitable amount of bamboo vinegar was discussed,so as to provide some references for improving germination conditions of fluecured tobacco seeds and the quality of tobacco seedlings,and reducing diseases and insect pests.

Materials and Methods

Experimental materials

The experimental varieties of tobacco were Yunyan 97,Guiyan 2,Jiucaiping 2 and Bina 1,the seeds were provided by Weining Branch Technique Center of Bijie Tobacco Company.Bamboo vinegar was produced by Zhejiang Suichang Wenzhao Bamboo-charcoal Co.,Ltd.,its density was 1.006 g/ml,pH was 2.62 and the content of acetic acid was 4.54%.

Experimental design

Effects of bamboo vinegar on seed germination of different flue-cured tobacco varietiesThe experiment was carried out in Organic Technology Research Center of China Agricultural University during February 10-25 in 2014.Culture dish filter paper method was used for seed germination experiment.Seeds of flue-cured tobacco with uniform plumpness and same size were chosen to sterilize by 75%ethyl alcohol for 2 min,then they were cleaned by sterile water for 3-4 times(the operation process was carried out on clean bench).The sterilized seeds were separately put on the culture dishes which had two layers of filter paper(9 cm diameter)for germinating,and each dish had 30 seeds,then they were separately added 5 ml bamboo vinegar aqueous solution with different dilution times (400,200,100 and 50 times),the control(CK)group was added 5 ml distilled water,after that,they were put into manual climatic box,thereby temperature shift for germinating was adopted(darkness:20℃and 12 h;illumination:30℃and 12 h,the illumination intensity was 4 000 lx),and the corresponding bamboo vinegar and distilled water were supplemented during the period,all these had three repetitions.Taking the breakthrough of testa with 2 mm for embryos as the standard,the germination number of seeds was observed and recorded every day.At the 14thday of seed germination,25 seedlings were chosen randomly,and vernier caliper was used to measure the overall length and root length of germinated seeds,thereby the stem length was calculated,then the fresh weight of the full stand was weighed,the germination rate,germination index (GI)and vigor index(VI)of seeds were counted,and the average value of the three repetitions was got.

In the formulas,nwas the number of germinative seeds at the final period of germination,Nwas total number of experimental seeds.

In the formula,Gtwas the number of germinative seeds at the tth day,Dwas germination days(d).

In the formula,Swas overall length of seedlings(cm).

Effects ofbamboo vinegaron seedling growth of Yunyan 97The experiment was carried out in seedling greenhouse of Heishitou Town,Weining County,Guizhou Province from February 26 to April 26 in 2014.Based on the results of seed germination experiment,the seeds of Yunyan 97 were chosen as experimental materials,four different concentrations of bamboo vinegar dilution(50,100,200 and 400 times)were designed and the same amount of clean water was used as CK,there were three repetitions and 15 seedling-raising plates,the plates were randomly put in the same nursery pond,differenttreatments were separated by dummy plates to avoid the interference generated by spraying medicaments.20 d after sowing,bamboo vinegar was sprayed on the leaves ofseedlings,then sprayed once every 10 d,and there were totally three sprays.Each spray was carried out in the afternoon of a fine day,and the bamboo vinegar was sprayed evenly until water drops on the leaf occurred.10 d after the 3rdspray,five-spot(removed the outermost tobacco seedlings)was adopted for taking out tobacco seedlings from seedling holes,15 tobacco seedlings were taken out for each treatment,then such agronomic traits as plant height,stem girth,the maximum leaf length and width,effective number of leaves were measured according to YC/T142-1998 method;after that,tobacco seedlings were put in 105℃for 15 min and dried in 60℃,then they were weighed and the weight of dry matter was calculated; drainage method(Li Yueet al.,2013)was used for measuringrootvolume;SPAD chlorophyll meter was used for measuring chlorophyll content.The result of data was the average value of 15 tobacco seedlings.

Statistical analysis

Microsoft Excel 2003 and SPSS 17.0 were respectively adopted for chart drawing and one-way analysis of variance,and Tukey s-b(K)method was used.

Results and Analyses

Effects of different bamboo vinegar treatments on seed germination rate of different flue-cured tobacco varieties

Germination rate of seedsAs shown in Table 1,seed germination rate of different flue-cured tobacco varieties decreased with the increase of bamboo vinegar concentration.Under the treatment of 50 times dilution,excessive concentration ofbamboo vinegarcaused the ungerminated seeds of different flue-cured tobacco varieties;under the treatment of 100 times dilution,the seeds of Guiyan No.2,Jiucaiping No.2 and Bina No.1 all did not burgeon.The differences on seed germination rates between Yunyan 97 and Guiyan No.2 under different bamboo vinegar treatments reached significant level,moreover,the germination rates of the above two were obviously less than those of CK.Seed germination rates of Jiucaiping No.2 and Bina No.1 under the treatment of 400 times dilution respectively reached 77.78%and 86.67%,and compared with corresponding CK,the differences were not significant,however,compared with the treatment of 200 times dilution,the two respectively increased by 42.87%and 36.85%,thus reached significant level.

Germination index of seedsAs seen in Table 2,all flue-cured tobacco varieties under the treatment of 50 times dilution did not bourgeon,so the germination index could not be counted.In 100-400 times dilution,seed germination index of different fluecured tobacco varieties increased with the decrease of bamboo vinegar concentration,and the differences among different treatments reached significant level.The germination indexes of Yunyan 97,Guiyan No.2 and Jiucaiping No.2 under the treatment of 400 times dilution were slightly lower than that of CK,and the differences were not significant,moreover,these under the treatments of 100 and 200 times dilution were obviously lower than that of CK,which effectively inhibited the germination of seeds.The germination indexes of Bina No.1 under the treatments of 200-400 times dilution were all significantly lower than that of CK.

Vigor index of seedsAs shown in Table 3,in accordance with germination rate and germination index,there was not vigor statistics of all flue-cured tobacco varieties under the treatment of 50 times dilution. Except Honghuadajinyuan,the seed vigor index of the other flue-cured tobacco varieties from low to high was as follows:100 times＜200 times＜400 times＜CK,and the differences among different treatments were significant,indicating that bamboo vinegar with high concentration could significantly inhibit the vigor of seeds.

The whole length of seedlings

From Table 4,it was drawn that the whole length of seedlings under different bamboo vinegartr eatments shortened gradually with the increase of bamboo vinegar concentration,moreover,they were all shorter than CK.The whole length of seedlings of Yunyan 97 under the treatment of 400 times dilution had not significant differences compared with corresponding CK,however,they were both significantly longer than that under the treatments of 200 and 100 times dilution and respectively increased by 1.84 and 3.43 times.The whole lengths of seedlings of Guiyan No.2 and Jiucaiping No.2 under the treatments of 400 and 200 times dilution were significantly shorter than those of corresponding CK,which were respectively 0.47 and 1.08 times,0.29 and 0.56 times,respectively.The whole length of seedlings of Bina No.1 under the treatment of 400 times dilution was shorter than that of CK,but the difference was not significant,while that under the treatment of 200 times dilutionwas significantly shorter than that of CK and reached 0.52 times.

Table 1 Effects of different bamboo vinegar treatments on seed germination rate of different flue-cured tobacco varieties

Table 2 Effects of different bamboo vinegar treatments on germination index(GI)of different flue-cured tobacco varieties

Root length of seedlingsFrom Table 5,it was inferred that root length of seedlings of different flue-cured tobacco varieties under different bamboo vinegar treatments was significantly shorter than that of CK except Yunyan 97 and Jiucaiping No.2 under the treatment of 400 times dilution.Root length of seedlings of Yunyan 97 under the treatment of 400 times dilution was 11.728 mm,which was longer than that under the treatments of 200 and 100 times dilution by 2.70 and 7.24 times.Rootlengths of seedlings of Guiyan No.2,Jiucaiping No.2 and Bina No.1 under the treatment of 400 times dilution respectively increased by 55.62%,35.00%and 16.38%compared with that under the treatment of 200 times dilution,but the differences were not significant.

Stem length of seedlingsFrom Table 6,it was concluded that the stem growth of seedlings was inhibited to varying degrees with the increase of bamboo vinegarconcentration;the stem length of seedlings of different flue-cured tobacco varieties under different bamboo vinegartr eatments from low to high was as follows:100 times＜200 times＜CK＜400 times.The treatment of 400 times dilution could significantly promote the growth of stem length of seedlings of Yunyan 97 and Bina No.1,which respectively significantly improved by 31.44%and 86.63%compared with CK,while the treatments of 200 and 100 times dilution had notobvious differences compared with CK.The stem length of seedlings of Guiyan No.2 and Jiucaiping No.2 underdifferenttreatments had not significant differences.

Fresh weight of seedlingsFrom Table 7,it was shown that the fresh weight of seedlings of different fluecured tobacco varieties under different bamboo vinegar treatments had not obvious rule.Under the treatment of 100 times,the seeds of all flue-cured tobacco varieties did not burgeon,or the growth vigour of seedlings was poor,thus the fresh weight can not be measured. The fresh weight of seedlings of Yunyan 97 under the treatment of 400 times was the heaviest,namely 3.057 mg,which significantly increased by 120.72%compared with that under the treatment of 200 times,but the difference with CK was not significant.The fresh weight of Guiyan No.2 and Jiucaiping No.2 under different treatments was as follows:200 times＜400 times＜CK,and the differences of them among different treatments were significant except Guiyan No.2 under the treatments of 400 and 200 times.The fresh weight of Bina No.1 under the treatment of 400 times respectively increased by 64.58%and 82.03%compared with that of CK and 200 times treatment.

Effects of different bamboo vinegar treatments on seedling growth of Yunyan 97

From Table 8,it was shown thatcompared with CK,spraying different concentrations of bamboo vinegar on Yunyan 97 at the seedling stage can promote the growth of seedlings to varying degrees,and most indexes of each treatment increased to varying degrees except the stem girth under the treatment of 200 times,the maximum leaf length under the treatment of 50 times and the chlorophyll content of leaves under the treatment of 100 times.

Table 3 Effects of different bamboo vinegar treatments on seed vigor index(VI)of different flue-cured tobacco varieties

Table 4 Effects of different bamboo vinegar treatments on the whole length of seedings of different flue-cured tobacco varieties mm

Table 5 Effects of different bamboo vinegar treatments on root length of seedlings of different flue-cured tobacco varieties mm

Table 6 Effects of different bamboo vinegar treatments on stem length of seedlings of different flue-cured tobacco varieties mm

Table 7 Effects of different bamboo vinegar treatments on fresh weight of seedlings of different flue-cured tobacco varieties mg

Table 8 Effects of different bamboo vinegar treatments on seedling growth of Yunyan 97

With the increase of bamboo vinegar concentration,the plant height,stem girth and the maximum leaf length of seedlings of Yunyan 97 showed the fluctuation variation trend of decline—rise—decline;the chlorophyll content of leaves showed the variation trend of rise—decline—rise;effective number of leaves rose at first,went down then,and finally maintained stable;the weight of dry matter decreased continuously.In all bamboo vinegar treatments,the plant height was the highest under the treatment of 100 times,namely 3.36 cm,which was significantly higher than that of CK and the other treatments.Each bamboo vinegar treatment had not significant effect on stem girth and weight of dry matter of tobacco seedlings,but they both were the highest under the treatment of 400 times,which were 1.82 cm and 0.262 g,respectively.Under the treatment of 100 times bamboo vinegar,the maximum leaf length of Yunyan 97 was the longest,namely 10.14 cm,which was significantly longer than that of 50 times treatment;under the treatment of 400 times bamboo vinegar,the maximum leaf width of Yunyan 97 reached 5.74 cm,which was obviously greater than that of CK and 50 times treatment.Except the treatment of 50 times bamboo vinegar,the root volume of all the other treatments was significantly greater than that of CK,in which that under the treatment of 100 times bamboo vinegar was the greatest,namely 0.92 ml.Effective numberofleaves and chlorophyll content under the treatment of 200 times were slightly higher than those of the other treatments,and there were not significant differences among different treatments.

Discussions

Effects of bamboo vinegar on seed germination and seedling growth of different flue-cured tobacco varieties

In recent years,many scholars did a large number of studies on the application effect of bamboo vinegar and pyroligneous acid on various kinds of crops,moreover,obtained good results.Muet al.(2003)and Liu Yanmeiet al.(2012)showed that bamboo wood fermentation broth with high concentration could inhibit the germinating ability of seeds.The results of this study showed that with the increase of bamboo vinegar concentration,the germination rate,germination index and seed vigor of almost all seeds of different flue-cured tobacco varieties decreased continuously,in which the seeds under the treatment of 50 times bamboo vinegar did not burgeon,which was similar to the results of previous studies.Muet al.(2004)demonstrated that bamboo vinegar with higher concentration had inhibiting effect on the root growth of seedlings,while the bamboo vinegar with low concentration had promoting effect.The results ofthis study showed that under the treatment of 400 times,only the overall length of Yunyan 97 and Bina No.1 had not significant differences compared with that of CK,and that of the other varieties showed decreasing trend to varying degrees, moreover, the seedling growth was inhibited significantly with the increase of bamboo vinegar concentration.

Effects ofbamboo vinegaron seedling growth of Yunyan 97

Spraying different concentrations of vinegar liquid had different effects on crop growth.Wu Xuanet al.(2009)showed that spraying low concentrations of bamboo vinegar(100 times and 300 times)could promote the growth of cucumber,while high concentration (50 times)would inhibit the growth of cucumber.Wang Xiaodonget al.(2009)studied the effects of straw vinegar on the seedling growth of teaoil tree,and held that the effect of 300 times straw vinegar on the seedling growth of tea-oil tree was the best.On the whole,the results of this experiment were similar with previous studies,that was,20 d after seed germination of Yunyan 97,such indexes as plant height,the maximum leaf lenth and width,weight of dry matter and root volume of tobacco seedlings under the treatments of 100-400 times bamboo vinegar were improved obviously,while these under the treatment of 50 times bamboo vinegar were slightly higher than CK or decreased,demonstrating that spraying low concentrations(100-400 times)of bamboo vinegaratthe seedling stage could effectively promote the growth and development of tobacco seedlings,but the effect of high concentrations of bamboo vinegar(such as 50 times)was not good.Therefore,the application effects ofbamboo vinegar before seed germination should be further studied in future,moreover,the method ofpouring bamboo vinegar into matrixes can be adopted,so as to verify its effect on regulating and controlling the microecologicalenvironment of matrix,thereby affecting the growth of root and improving the quality of tobacco seedlings.

Conclusions

The results of this experiment showed that supplying different concentrations of bamboo vinegar on the culture dish could inhibit seed germination and seedling growth of fluecured tobacco;during the process of float breeding,spraying 100-400 times bamboo vinegar solution to leaves could promote the seedling growth and improve the quality of Yunyan 97 to varying degrees.

[1]FANGY(方楷),YANGGY(杨光耀),YANG QP(杨清培).Progress in application of vinegar(竹醋液应用研究进展)[J].Forestry Science and Technology(林业科技开发),2007,21(6):1-4.

[2]ZHANG WB(张文标),YE LM(叶良明),LIU L(刘力),et al.Analysis on component of bamboo vinegar(竹醋液的组分分析)[J].Journal of Bamboo Research(竹子研究汇刊),2001,20(4):72-76.

[3]SULAIMAN O,MURPHY RJ,HASHIM R,et al.The inhibition of microbial growth by bamboo vinegar[J].Journal of Bamboo and Rattan,2005,4(1):71-80.

[4]LI XR(李小荣),WU QC(吴全聪),LIU ZL(刘志龙),et al.Synergistic effect of bamboo vinegar on several kinds of insecticides(竹醋液对几种杀虫剂的增效作用)[J].Zhejiang Agricultural Sciences(浙江农业科学),2005(2):144-146.

[5]ZHANG WB(张文标),ZHONG TL(钟泰林),ZHANG H(张宏),et al.Effects of bamboo vinegar on seed germination and seedling growth of Yumai 18(竹醋液对豫麦18种子发芽及生长的影响)[C].//Chinese Society of Forestry(中国林学会).First Bamboo Industry Conference Proceedings of Chinese Society of Forestry(中国林学会首届竹业学术大会论文集),2004.

[6]WEI Q(韦强),DU XG(杜相革),QU ZH(曲再红).Effect of treatments with bamboo vinegar on growth of cucumber(竹醋液对黄瓜生长的影响)[J].Chinese Agricultural Science Bulletin(中国农学通报),2006a,22(7):411-414.

[7]YANG H(杨晖).The delayed effects of bamboo vinegar on senescence of cut tulip(竹醋液对郁金香切花衰老的延缓作用)[J].Zhejiang Agricultural Sciences(浙江农业科学),2005(4):264-265.

[8]ZHOU ZC(周志成),XIAO QM(肖启明),ZENG AP(曾爱平),et al.Tobacco pest and disease control(烟草病虫害及其防治)[M].Beijing:Chinese Agriculture Press(北京:中国农业出版社),2009:22-32.

[9]HU CS(胡春水),LIANG WB(梁文斌),LI JA(李建安).EM,research Zhucu pollution in forest cultivation application(EM、竹醋在经济林无公害栽培中的应用研究)[J].Jiangxi Forestry Science and Technology(江西林业科技),2000(1):4-6.

[10]WEI Q(韦强),DU XG(杜相革),HUANG MQ(黄漫青).et al.Study on the effect of bamboo vinegar on controlling downy mildew of cucumber(竹醋液对黄瓜霜霉病防治效果的研究)[J].Chinese Agricultural Science Bulletin(中国农学通报),2006b,22(6):330-332.

[11]SUN JH(孙剑华),CHEN YN(陈永宁),SHEN XK(沈晓昆).Effect trial of pepper Juanyeling combining bamboo vinegar on controlling pepper virus disease(辣椒卷叶灵加竹醋液防治辣椒病毒病药效试验)[J].Agricultural Equipment and Technology(农业装备技术),2006,32(1):35.

[12]ZHANG L(张琳),DONG L(董琳),WANG JC(王甲辰),et al.Influence of wood vinegar as leaves fertilizer on growth and quality of tomato(木醋液作为叶面施用对西红柿长势及品质的影响)[J].Beijing Agriculture(北京农业),2010(30):58-62.

[13]YAO Y(姚远),CHEN ZZ(陈正洲),HONG K(洪科),et al.Effects of bamboo vinegar with different concentrations on the growth and development of tomato andMyzus persicae(竹醋液对番茄生长发育及桃蚜的影响)[J].Journal of Anhui Agricultural University(安徽农业大学学报),2012,39(4):646-650.

[14]LI Y(李 玥),SHEN H(沈宏),JIA ZH(贾志红),et al.Effects of a specific tobacco root promoting agent on root growth of flue-cured tobacco(一种烟草专用促根剂对烤烟根系生长的影响)[J].Chinese Tobacco Science(中国烟草科学),2013,34(3):10-15.

[15]MU J,UEHARA T,FURUNO T.Effect of bamboo vinegar on regulation of germination and radicle growth of seed plants[J].Journal of Wood Science,2003,49(3):262-270.

[16]LIU YM(刘艳梅),WANG XX(王晓旭),WANG XF(汪晓峰),et al.Effect of pyroligneous acids on seed germination ofProtea cynaroides(木醋液对帝王花种子萌发的影响)[J].Journal of Zhejiang Forestry Science and Technology(浙江林业科技),2002,32(4):51-53.

[17]MU J,UEHARA T,FURUNO T.Effect of bamboo vinegar on regulation of germination and radicle growth of seed plants II:compositon of mose bamboo vinegar at different collection temperature and its effects[J].Journal of Wood Science,2004,50(5):470-476.

[18]WU X(吴暄),ZONG LG(宗良纲),LIU X(刘新),et al.Growth and pests control in organic cucumber production(竹醋液对有机栽培黄瓜生长的影响及其对蚜虫的防治效果)[J].Chinese Journal of Biological Control(中国生物防治),2009,25(4):309-311.

[19]WANG XD(王小东),ZHOU WC(周文才),WANG XP(王小平).Effects of straw vinegar on growth ofCamellia oleiferaseedlings(秸秆醋液对油茶幼苗生长的影响)[J].Jiangxi Forestry Science and Technology(江西林业科技),2009(5):10-11.