Research On Tissue Culture And Changes Of Endogenous Hormones In Subeculture Of Chinese Fir(Cunninghamia Lanceolata)

Chinese fir,Cunninghamia lanceolata(Lamb.)Hook,is a unique specie of fast-growing timber in our country,which has the features of fast growth,high yield,good material and wide application.Consequently,it has become one of the most important material goods planted in south China.With the rapid development of biological engineering technology in recent years,especially in plant tissue culture technology and its rapid propagation system,domestic and foreign scholars have successively carried out research on Chinese fir in biotechnology and achieved some results.The way using tissue culture technology to Chinese fir clones were studied,which effectively solve some defects of traditional breeding way.As a meansof commercial production,tissue culture technology can meet the huge demand of Chinese fir seedlings in market and also is an important way to preserve rare provenance and clone.With the continuous improvement of endogenous hormone detection means,domestic and foreign scientists have studied the change of endogenous hormones of some plants in the process of plant tissue culture in recent years.Chinese fir clones of high propagation coefficient were researched and screened out,which provides theory basis for the similarities and differences of different Chinese fir clones in vitro culture and provides scientific basis and technical support for improving the rapid propagation technology of Chinese fir tissue culture,boosting factory production and establishing genetic transformation system of Chinese fir.This will have important practical significance of forestry production in southern forest area.In view of this,this thesis uses FS09 and YKC05 two Chinese fir clones as experimental material,which has established a stable rapid propagation system of Chinese fir in vitro,completed the proliferation of fir tube seedlings and the filtering of rooting medium and explored the effects of organic additives on proliferation and on endogenous hormones of Chinese fir.The main results are as follows:1?The best formulas for proliferating and cultivating of Chinese fir clones FS09 and YKC05 are:MS+0.3 mg/L 6-BA+0.2 mg/L IBA + 40 g/L Sucrose + 5.4 g/L Agar +pH 5.8.Their highest proliferation coefficients are 11.38 and 9.6 respectively.The best formulas for growing of Chinese fir clones FS09 and YKC05 are:MS+0.2 mg/L 6-BA+0.2 mg/L IBA+40 g/L Sucrose + 5.4 g/L Agar + pH 5.8.The highest maximum values are 5.37 cm and 6.6 cm.2.The formulas of rooting medium for clones FS09 is:1/4 MS + 0.12 mg/L IBA +0.09 mg/L NAA + 10%potato + 0.25 g/L AC.The formulas of rooting medium for clones YKC05 is:1/4 MS + 0.12 mg/L IBA + 0.06 mg/L NAA + 10%potato + 0.25 g/L AC.Organic additives and activated carbon togeter have significant effects on rooting rate.IBA has very significant effect on rooting rate.After adding 0.12 mg/L IBA,rooting effect is best.3.When the concentration of potato is 10%,the proliferation coefficient and plant height of clone FS09 reaches to maximum,13.19 and 7.09 cm respectively.When the concentration of coconut juice is 20%,the proliferation coefficient and plant height of clone YKC05 reaches to maximum,13.35 and 7.41 cm respectively.This shows the coconut milk has obvious role in promoting proliferation of clone.4.The effects of different organic additives on the endogenous hormone levels of Chinese fir clones FS09(1)Change trend of ZT is complex,which is rising and falling after falling to rise again.In the training process of adding potato,the ZT content in different stages always guarantees minimum.In the training process of adding banana and peptone,ZT hormones are greater than controls,which shows there is analogue of endogenous hormone ZT existing in Banana and peptone.The analogue may participate in the growth of the somaclone and promote cell division and differentiation.(2)After different additives processing,IAA content reduces first then rises and reduces again when 40th day.The peak values of banana and peptone increase by 29%and 29%compared with the peak value of contrast,which has significant difference.The peak values of experimental group and contrast appear at the same time.Compared with the peak value of contrast,the peak values of coconut milk and potato reduce by 13.43%and 13.43%respectively,which appears 10 days earlier than contrast.(3)The content of ABA reduces after rising first and changes little in different treatment times.The highest content of ABA is 39.14 ?g/g when adding coconut milk in 30th day,which significantly increases by 35.72%compared with the control.The content of ABA is 24.81 ?g/g when adding peptone in 30th day,which reduces by 13.96%compared with the control.After adding coconut juice,the content of ABA increases rapidly in 20th to 30th day and reduces rapidly in 30th to 40th day.(4)The trend of GA3 content is increasing after reducing first.And the content changes largely with different additives.Add the coconut milk,the processing of GA3 peak of 19.27 ?g/g,controlled peak increased 2.37 times,significant difference.And add the processing of organic additives,after 30th day were compared with high concentration.5.The effects of different organic additives on the endogenous hormone levels of Chinese fir clones YKC05(1)The total trend of content change of ZT is complex and the content changes largely with different additives.Compared with the peak value of control group,the processing peak value of adding bananas and peptone don't have much difference.compared with control group,the content of ZT in coconut milk group in 50th day is and is reduced by 48.5%.(2)The change of IAA content reduces first and increases then and declines again in 30th day.Furthermore,with the extension of processing time,the content has a tendency to significantly higher.Compared with control group,its content in processing group of adding banana,peptone and potato significantly increases,content in coconut milk is reduced by 22.92%and content in peptone is growing rapidly between 20th d and 30th d.(3)The change of ABA content increases first and reduces then and the effects of different organic additives on the content of ABA aren,t significant.The ABA content of potato group increases to the highest value in 20th d,up to 30.04?g/g and is increased by 4.49%,compared with control group.(4)The change of GA3 content reduces first and increases then,affected by different additives significantly.There is a significant difference that the GA3 content of potato group increases to the highest value,up to 31.23 ?g/g and is increased by 73.08%,compared with control group.The GA3 contents in groups of adding coconut milk,peptone and banana 30th d are higher than it in control group.