Study On Excellent Breed Screening And Quality Evaluation Of Amomum Villosum Lour

Amomum Villosum, also known as Fructus Amomi,a geo-authentic crude drug in Yangchun district of Guangdong province, is derived from the dried ripe fruit of Amomum Villosum Lour, from Zingiberaceae plants.The quality of Amomum villosum natived to Panlong and Chunwan district is best, especially in Jinhuakeng. Amomum Villosum is one of four famous southern medicines in China, which has function of eliminating damp, improving appetite, warming spleen, checking diarrhea and preventing abortion. It is commonly used in treating intestines or stomach diseases and becomes one of the raw materials of Chinese patent medicine such as "Xiangshaliujunwan","Xiangshalizhongwan".Because the fruit fragrance of Amomum Villosum is very rich and nice, the taste is a little bitter,sour and spicy, Amomum Villosumhas been widely used in producing health food, which becomes more and more popular among consumers from home and abroad.In order to solve national pollination problem and improve yield of Amomum Villosum, we carried out studies on excellent breed screening and quality evaluation of Amomum Villosum. On the basis study of germplasm resources investigation of Amomum Villosum in Yangchun district, we analyzed and identified the germplasm of four different cultivated forms of Amomum Villosum in Yangchun so as to sift the most excellent germplasm. Secondly, we also have bred several generations of the hybrid, contrasting spring No.1-F4(the forth generations of hybrid) and its parents (Longfruit No.2as the female parent, A. longiligulare T. L. Wu as the male parent) from the aspects of output, molecular characteristics and water-soluble polysaccharide content.Moreover, we have conducted backcross inheritance experiment using Longfruit No.2as female parent, Spring No.1-F4as male parent to acquire Spring No.2. Finally, we compared the morphological characteristics between Spring No.2and its parents. In conclusion, this paper sift and bred an excellent germplasm of Amomum Villosum to lay the foundations for solving natural pollination problems, breeding high quality products, expanding resources of Amomum Villosum.The mainly content was reported as follow.Chapter One:Literature Reviewwe have summarized systematically the study of Amomum villosum based on a great deal of investigating and studying on relational information, including the previous studies on textual research, approbation, chemical component, pharmological study,quality evaluation, cultivation research and breeding research.Chapter Two:Experimental ResearchSection one:the screening and comparative research on different cultivars of Amomum Villosum.We compared and identified four cultivars of Amomum villosum from growth characteristics, distribution, infrared spectrum characteristics and output in Yangchun district in order to select a high quality germplasm with the strongest disease-resistant rate and highest pollination rate.Part one:the distribution and growth characteristics of four cultivars of Amomum villosumObserving the distribution and growth characteristics of Amomum villosum such as florescence, lower-type structures, pollen characteristics and fruit characteristic on a regular basis,the result have shown that Longfruit No.2is the last coming into blossom than the others,which is conducive to insect pollination, so its natural pollination rate, seed setting rate and fruit set percentage are higher than the others. In addition, the resistance of Longfruit No.2is also stronger than others. Therefore, Longfruit No.2is suitable for cultivation and also widely grown in Yangchun,Guangdong province.Part two:the comparative study of infrared emission features of four cultivars of Amomum VillosumThrough the comparative analysis and research of these charts of infrared spectrophotometry features of the chemical constituents of four cultivars of Amomum Villosum with the methods of Fourier Transform Infrared Spectroscopy (FT-IR) and Multi-order derivative, the results have shown that four cultivars of Amomum Villosum have similar infrared absorption in the wan band between750-1800cm-1and2700-3500cm-1. Besides, these graphs have also shown no significant difference after fourth derivative.It has illustrated that the changing ecological environment and different cultivated methods have no effect on chemical structure of four cultivars of Amomum Villosum, the four cultivars still are belong to the same strain.Part three:the comparative analysis of output of four cultivars of Amomum VillosumA field experiment was conducted to study the difference between setting rate and yield of four cultivars of Amomum Villosum in the condition of natural pollination and artificial pollination. The results have shown that both the setting rate and yield of four cultivars of Amomum Villosum in artificial pollination are higher than that in natural pollination. In addition, the setting rate and yield of Longfruit No.2are the highest in two conditions. It is concluded that Longfruit No.2is the most excellent cultivar.Section two:the comparative study of Spr ing No.1which is the hybr id of Amomum Villosum through oriented cultivation and its parentA new hybrid (Spring No.1) was bred by Professor Zhang using Longfruit No.2as female parent and A. longiligulare T. L. Wu as male parent. On this basis, we have gained Spring No.1-F4by means of oriented cultivation.Moreover, we evaluated the quality of Spring No.1-F4through comparative analysi s from yield, molecular characterization and water-soluble polysaccharide contents between Spring No.1-F4and its parent (female parent or male parent).Part one:the oriented cultivation of the hybrid (Spring No.1-F4) of Amomum VillosumSowing the seeds of Spring No.1, we got Spring No.1-F4after self-crossing.Part two:comparative analysis on yield between Spring No.1-F4and its parentA field experiment was conducted to study the yield difference between Spring No.1-F4and Longfruit No.2in the condition of natural pollination and artificial pollination. The results have shown that the yield of Spring No.1-F4is far higher than Longfruit No.2in the two conditions, and the yield of Spring No.1-F4in the condition of natural pollination is similar with the yield of Longfruit No.2in the condition of artificial pollination, which also proved that Spring No.1-F4can gain the ideal output without artificial pollination. Part three:the comparative analysis of molecular properties between Spring No.1-F4and its parentComparative analysis on peaks of ITS gene sequence between Spring No.1-F4and its parent was conducted. The results have shown that the ITS sequence of Spring No.1-F4have nine degenerate bases, all which are the allelic superposition of its female parent and its male parent, these changes in ITS sequence conform to the Law of Mendelian Genetics, the feature proved that Spring No.1-F4is the hybrid of Longfruit No.2and A. longiligulare T. L. Wu.Part four:comparison analysis on water-soluble polysaccharide contents between Spring No.1-F4and its female parentDetermining water-soluble polysaccharide contents of Spring No.1-F4and Longfruit No.2by using ultrasonic technology extraction, alcohol deposit and Sevag deproteinization. The results have shown that the contents of water-soluble polysaccharide in Spring No.1-F4are higher than that in Longfruit No.2, which indicates that the pharmacology of Spring No.1-F4may be better than Longfruit No.2.Section Three:the backcross inheritance experiment of Amomum VillosumAlthough Spring No.1-F4has advantages of high yield and strong resistance,the poor fruit taste and high plant height hinder the development of the hybrid. Therefore, we have conducted backcross inheritance experiment using Longfruit No.2as female parent, Spring No.1-F4as male parent in order to gain a more excellent backcross which inherited good qualities from its mother parent.Part one:the method of backcross breedingSpring No.1-F4was taken as male parent and Amoinum Villosum cultivated type Longfruit No.2was taken as female parent. Bagging the female parent flower when it was in the pre-flowering stage and manual emasculation was carried out in blooming period. Pollen of male parent was scraped to be pollenized into stigma of the flower of treated female parent and the flower of female parent was protected by bagging again.After hybridization of female parent, other undeveloped flowers of the inflorescence were removed. A bag protector was removed and management was reinforced2to3days later. The mature fruits were pack up after scorching than we gained Spring No.2after sowing the seeds in September. The young plants were transplanted and watered carefully to the field in August next year. Part two:the comparative analysis of morphological characteristics between the backcross and its parentsComparing the morphological characteristics such as plant height, spear thickness and leaf length/width of Spring No.2with Longfruit No.2and Spring No.1-F4. The results have showed that plant height and spear thickness of Spring No.2have inherited its female parent, leaf length/with of Spring No.2have inherited its male parent.GonelusionThe study of system selection combining with cross breeding was carried on with the purpose of solving natural pollination problem and improving output of Amomum Villosum. On the basic study of germplasm resource investigation of Amomum Villosum in Yangchun district (the genuine producing areas of Amomum Villosum in China), we analyzed and identified growth characteristics and quality of four different cultivated forms of Amomum Villosum in Yangchun district so as to screen the most excellent germplasm. Secondly,we have bred several generations of the hybrid,then comparing the hybrid (Spring No.1-F4) with its parents (Longfruit No.2as the female parent, A. longiligulare T. L. Wu as the male parent) from output, molecular characteristics and water-soluble polysaccharide content. We also have conducted backcross inheritance experiment using Longfruit No.2as female parent, Spring No.1-F4as male parent to acquire Spring No.2. Finally, we compared the morphological characteristics between Spring No.2and its parents. In conclusion, this paper sift and bred an excellent germplasm of Amomum Villosum to lay the foundations for solving natural pollination problems, breeding high quality products, expanding resources of Amomum Villosum.