| Agarwood is one of the most famous medicinal and perfume resources in the world.In many Asian countries,the industry of agarwood is both traditional and new industry.The large-scale application of high-quality agarwood germplasm resources and efficient agarwood production technology is great significance to the sustainable development of agarwood industry.The Agar-WIT has been widely used in cultivated Aquilaria forests and can produced agarwood statively,but the quality change of agarwood induced by the technology is not clear yet.High quality Aquilaria germplasm resources have been the focus of agarwood industry in recent years,but the high quality germplasm not being able to propagate sexually,and there are technical barriers to large-scale breeding.In this paper,we studied the quality of agarwood produced by Agar-WIT and the tissue culture technology of A.sinensis.the change rule of agarwood quality after the implementation of Agar-WIT was basically defined,and the technical system in the tissue culture of A.sinensis was optimized and improved.The obtained results as follows.1.Quality Monitoring of Agarwood Produced by Agar-WIT.(1)The research of agarwood induced for one month:This study explored the change characteristics of the macroscopical and microscopic properties,percentage of agarwood,agarotetrol content,Some sesquiterpenoidsn and aromatic compounds of agarwood within one month.It can be concluded that the Agar-WIT can quickly induce the formation of agarwood.The results showed that the agarwood layer structure can be observed macroscopically within 4-5 days after the treatment,and the resin contained in the Interxylary phloem,vessel and rays of the agarwood can be observed by microscopy.The average agarwood rate within one month is 8.04%.The agarotetrol is synthesized within 1 week of induction,and the content increased continuously within one month.Some aromatic compounds begin to synthesize and accumulate after 1 week of induction,and some sesquiterpenoids start to synthesize and accumulate after 2 weeks of induction.(2)The research of agarwood induced for one year:In this study,the percentage of agarwood,barrier layer,extract content,agarotetrol content and characteristic chromatogram of agarwood were examined.It is clear that the Agar-WIT can efficiently induce the formation and accumulation of agarwood and is suitable for large-scale production of agarwood in cultivated agarwood forests.The results showed that Agar-WIT can maintain a higher percentage of agarwood which could reach 10-18%.Alcohol-soluble extract content and agarotetrol showed a fluctuating cyclical change,but the overall trend is increasing.Two indices reached the peak for the first time in the 5th month and the 6th month,and the second peak in 11th month,respectively.The characteristic chromatogram of agarwood from 1-12 months is consistent with the reference medicinal material.After 4 months of Agar-WIT induction,the agarwood had met the Chinese pharmacopoeia standard.The barrier layer began to appear in the 4th month after the treatment.On.the basis of the overall high yield of agarwood,the 11th month and the 5th to 6th month should be the best harvest time.(3)Quality analysis of the Agarwood in various producing areas of domestic:Through the study of macroscopic properties,extract content,agarotetrol content and characteristic chromatogram of agarwood samples that have been processed for more than 6 months in Hainan,Guangdong,Guangxi,Yunnan and Fujian provinces and regions,it was found that the samples of Agarwood also meet the standards of the 2020 Chinese Pharmacopoeia.For samples from the same province,the alcohol soluble extractive contents and agarotetrol are generally positively correlated.2.Research on Tissue Culture and Rapid Propagation Technology of Aquilaria sinensis(Lour.)Spreng.(1)The study of explant disinfection:In this study,the stems of field branches were used as explants,and the effects of pretreatment methods,disinfection methods and sterilization agents on the survival of explants were research,and the best sterilization method for field branch explants was successfully obtained.The results showed that the explants collected in the field can be treated by conventional disinfection methods.In winter when the light is weak and the temperature is low,the explants are soaked in carbendazim solution for 3 minutes,rinsed in running water for 3 hours,and disinfected with 0.1%mercury liter solution for 6?8 minutes,which can effectively improve the survival rate of explants.In addition,adding 1 ml·L-1 preservatives(kasong)or 2?3 ml·L-1 phytocidin in the culture medium can also be used to sterilize stem explants taken to the field in winter.(2)The study of Multiple shoots induction:This study explored the effects of explant materials,basic media and plant growth regulators on the induction of clumping buds,and successfully obtained the best induction conditions for the clumping buds from the sterile seedlings and field branches.The results show:?Multiple shoots induction for seedlings was in Woody Plant Medium supplemented with 20 g·L-1 socruse,0.1 mg·L-1 6-Benzylaminopurine(6-BA)and 0.2 mg·L-1 naphthalene acetic acid(NAA).?The shoots and stems are used as explants in 1/2 MS+25 g·L-1 sucrose+2 mg·L-1 6-BA+0.01 mg·L-1 NAA medium,1/4 MS+20 g·L-1 sucros+2 mg·L-1 6-BA+0.5 mg·L-1 NAA medium,WPM+20 g·L-1 sucrose+1 mg·L-1 6-BA+0.01 mg·L-1 NAA medium can induce sprout buds.Then,cut the enlarged cluster of buds as a whole and transfer them to WPM+20 g·L-1 sucrose+0.1 mg·L-1 6-BA+0.2 mg·L-1 NAA medium,which can grow.(3)Research on Rooting:In this study,the rooting of cluster buds induced by two materials of sterile seedlings and field branches was carried out.According to the results of rooting induction found that adding 20?30 g·L-1 sucrose to the WPM medium and adding 0.1?0.2 mg·L-1 NAA can rooting the shoots induced by sterile shoots and explants from stem segments.In addition,the rooting of the sterile seedlings is easier,and the rooting of the cluster buds induced by the tender stems of the field shoots is more difficult. |
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