Resource Investigation And Genetic Diversity Analysis Of Chinese Fir King In China

Chinese fir is an endemic species in China, which has a lot of good characteristic such as rapid growth, high yield, good reproductive performance, et al. With the continuous progress of the forest rights reform in Fujian province, a large amount of natural forest including broad-leaved forest has been destroyed in order to plant the fir forest. Therefore,there is a sharp decline in the germplasm resources of natural fir forest. Scholars has researched Chinese fir much more, such as fir seed breeding,but most of the material is different fir seed source which is the allocation of second-generation seed orchard material. Therefore,the study on genetic variation of Chinese fir trees in China, Chinese fir center production area can provide an important evidence to reveal the distribution patterns of Chinese fir trees in China.Chinese fir king has experienced the natural ordeal for a long time,and during the process,it has record the changes of climate history,which made it be full of great scientific value. Yu Xin-tuo,etc. had investigated the growth condition of the20Chinese fir king in Fujian Province in1979,Huang Mu-sheng had investigated and compared the genetic diversity of the existing44different geographical provenances of Chinese fir king by ISSR molecular marker technique in2009,but there is no relevant reports about the resources distribution,particularly in relation to the number of Chinese fir king,growth conditions in other provinces and their genetic diversity is also scarce. For this reason, this study investigates the number of Chinese fir king, growth conditions and distributing,and compares the genetic diversity of the existing30different geographical provenances of Chinese fir king by SRAP molecular marker technique,and analyzes Eco-fir and Genetic variation of genetic about Chinese fir king,At the same time analyzes samples for genetic relationship,using genetic dissimilarity coefficient for different provenances,which revealed a different provenances Genetic variation of Chinese fir king and population distribution patterns, providing the molecular level evidences on,the range of Fujian fir Center production and population distribution patterns for a reasonable division of Chinese fir provenances,breeding programs to develop and germplasm resource management but also providing a scientific basis for Chinese fir seed source germplasm gene pool for the establishment and protection of ancient and famous trees The major findings are as follows:1、The comprehensive survey about Chinese fir king germplasm resources indicates that:Chinese fir king germplasm distribute in their natural growing areas in the provinces. Most of Chinese fir king grown in average annual temperature of17℃-20℃,1000-1500ml of rainfall,frost-free period of270days. Chinese fir king grow in different Provinces with incomplete information, such as missing age records and growth conditions (altitude,slope).2、The comprehensive survey about Chinese fir king germplasm resources indicates that:according to the standard about Chinese fir king, its diameter at breast height, tree height and plant volume respectively over1m,30m, and10m3, the old and big fir can be called Chinese fir king. Summaries of the Chinese fir king resources survey found that all the king’s growing Chinese fir is poor, tree height and diameter at breast height grow slowly and with little seed. Most of them grow in the hillside slope or roadside hobbled horse. They are protected by local village management. Some grow in the mountains with little management and less human disturbance.3、The DNA extraction of Chinese fir king has stabled. The method which is suited for Chinese fir king, the fir leaves should be sunk in liquid nitrogen and are ground for5-6times to ensure that which are ground into powder,sample in liquid nitrogen to prevent oxidation, using PW to extract DNA twice, the extracted DNA is suitable for SRAP-PCR.4、A valid reaction system suitable for the SRAP analysis about Chinese fir king was established (20μL.total reaction buffer),composed by80ng template DNA,200μmol/L dNTP,1.0umol/L premier,Mg2+2.0mM,2U Taq polymerase;The PCR parameter is:94℃pre-denature for5min,94℃for1min,40℃for1min72℃for1min,then5cycle;94℃for1min,50℃for1min,72℃for1min,then35cycle;finally extension at72℃for10min,the product of PCR was kept at4℃.5、Eighteen primers which are selected from one hundred and ten primers tested on the base of the number and frequency of polymorphisms are utilized in analyzing genetic diversity and relationship of Chinese fir king. They had154excellent amplification DNA.One hundred and fourty-four Polymorphic loci are amplified and the average percentage of polymorphic sites are93.51%.it showed rich polymorphism.6、According to Nei-Li method.it calculated that the genetic similarity range of32different Chinese fir king germplasm was between0.5195and0.9326. the average genetic similarity0.6893.4and15have the least genetic similarity0.5195.27and28have the largest genetic similarity0.9326.Description SRAP markers can effectively reveal that the genetic differentiation of different Chinese fir king provenances is at a high level.7、The results of the SRAP-PCR and UPGMA clustering analysis showed that the genetic diversity of Chinese fir king was not at a low level,the result of germplasm genetic distance clustering is quite different from the geographical distribution,which shows that not only geographical distance and differences in their environment different conditions but also the genetic basis of Chinese fir king may be the important factor of germplasm genetic variation. In the genetic similarity of0.72,32chinese fir king provenance are Divided into six groups, Jiangxi Yongxiu and Hunan Chengbu, are respectively independent groups.The third group is the North fir zones include:Anhui Maoshan, Anhui Xiuning, Hubei Jianshi three points; Guangdong Huaiji, Guangdong Pingyuan, Guangdong Shixing, Fujian Hua’an, Jiangxi Chongyi are in the same group;the fourth group is Fujian Pucheng and Hunan Changsha, the fifth group is divided into three categories:the first sub-category is the Yangshan, Guangdong;Xishui, Guizhou; Changshun, Guizhou;Changde, Hunan, Yucheng,Sichuan; and the second sub-category is Jianhe, Guizhou;Qingyuan, Zhejiang; Luding,Sichuan; third sub-category is Wuyuan,Jiangxi; Guangchang,Jiangxi;Maguan;Jinxiu,Guangxi; Rongshui, Guangxi; Zhenxiong, Yunnan;Taiwan; Yongti,Fujian; The sixth group is Yanping Fujian andMinqing Fujian.8、Study on17fir reference sample cluster, in the genetic similarity of0.66, germplasm samples were divided into three groups, the first group is Chengbu2,Hunan, the second group is Xiuning Anhui, Maoshan, Anhui, Huaiji,Guangdong;Shixing,Guangdong; Ping Yuan,Guangdong; Yangshan Guangdong; Luding Sichuan, Changde Hunan, Taiwan, Rongshui Guangxi; Chengbu Hunan; Jinxiu Guangxi;Chongyi, Jiangxi;Changsha, Hunan; Wuyuan Jiangxi,Minqing,Fujian.The clustering results showed that:17fir reference sample in the genetic similarity of0.66are divided into three groups. The first group is a independent group Chengbu2;in the genetic similarity of0.70the second group can be divided into two subcategories, the first sub-category is Xiuning Anhui Maoshan Anhui, Guangdong, and Huaiji Guangdong three reference samples, xiuning Anhui and Maoshan Anhui with higher genetic similarity; the second sub-PingYuan Guangdong, Shixing, Guangdong Huaiji Guangdong three references, all of them grow in northern Guangdong fir. in the genetic similarity of0.73,the third group can be divided into three sub-categories, the first sub-category is Luding Sichuan and Changde Hunan, the second sub-category is Taiwan, Rongshui Guangxi;Chengbu Hunan; Jinxiu Guangxi;Chongyi Jiangxi;The third sub-category is Changsha Hunan; Wuyuan, Jiangxi and Minqing Fujian reference.