VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE FACULTY OF AGRONOMY UNDERGRADUATE THESIS “GENETIC DIVERSITY OF THE PURPLE WAXY INBRED LINES REVEALED BY PHENOTYPE AND SSR MARKERS” STUDENT : NGUYEN THI THUY LINH CODE : 621691 CLASS : K62KHCTT SUPERVISOR : PROF. VU VAN LIET DEPARTMENT : PLANT GENETICS AND BREEDING HA NOI- 2022 COMMITMENT I pledge that the contents of this report are honestly conducted by me under the guidance of Prof. Vu Van Liet. I hereby undertake that all references in this document are quoted which are the name of the authors, name of research, time, location, and origin.
Hanoi, March 2022 Student Nguyen Thi Thuy Linh ACKNOWLEDGMENT During the implement this graduation thesis, I am not the only effort but also received a lot of support in many aspects of leaders, organizations, and individuals. First of all, I would like to express my sincere thanks to the dean of the Agronomy Faculty in general, my teachers in the Department of Plant Genetics and Breeding in particular who taught and created most conditions for students in the process of learning and performing this research. The project would not have been possible if it lacks support from the mentors of the teachers. Therefore, I would like to express my deep gratitude and respect to Prof.
Vu Van Liet has dedicatedly instructed, cared for, and helped me during the last time. Their ideas, inputs, and guidance were most helpful and have assisted me in making the research. Moreover, I also would like to thank the dean and all the teachers who have imparted knowledge to me, facilitating and supporting me during the implementation of the topic. This is a great opportunity I was able to practice my skills in class.
At the same time, it was good to help to give me more confidence in myself. Finally, I would like to send my sincere thanks to my family, friends, and relatives for encouraging, supporting, and helping me during the thesis. Hanoi, March 2022 Student Nguyen Thi Thuy Linh CONTENTS 090/10/1091) 002222.----- Ác Làn HT TH TT HH rệt 1 LIST OE TAB. ------ sàng HT HT HT HT HH V LIST OF FIGURES 111.
i LIST OF ACRONYMS.--- LH HH HH Hàng HH kiệt 1 PART I. ObJective and reQu1T€IT€TIS .---- + ¿+ +22 2E *kEéE#EEkrkrgvrrret 2 In 90. v9 Hàn nh HH HH ng nh ri 2 PART ¡001605 2. History original of the purple WaXY COFTA.
Waxy corn classification 2. Characteristics of waxy corn 2. Purple WaXY COFI. - ST TT TT HT HH TH nrhrêt 6 2.
- ch SH HT TH HH nh ro 6 2. Research and production of waxy corn in the WOrÌd. Genetic studies on tralfS i1 WAXY COFT.- 5< + xxx vs skkekrses 11 2. Inheritance of quality traits in waxy corn 2.
Genetics of purple waxy corn 2. Improved quality of waxy corn 2. Achievements in breeding anthocyanin-rich purple waxy corn. Situation of research and production of waxy corn in VIETNAM.
SSR markers and it function in plant breeding. Research on waxy corn gøenetic diversity anaÏySIS. Research on waxy corn genetic diversity analysis based on phenotypes and molecular markers In the WOTÏ(. Research on genetic variation based on phenotype and molecular Markers in i€ffiam.
---- 2+ S2 122211132311 32311 11 111 vn ven 27 PART II: MATERIALS AND METHOIDS .--- - - <5 S22 <2<ccsz 30 Ein son. Research place and tIIe.---- 5- + + + 23k HH HT Tri, 30 3.- Gà HH HT TH HH TH Hà HH TH TH Hà HH nh ng 3. ---- 5 + kg HH Hi, 30 3.2 Applied technicaÏl meaSUTS. Methods of moniforing_ eXp€TII€T(S.
DNA extraction and SSR analyS€S. PART IV: RESEARCH RESULTS AND DISCUSSION 4. Growth and development stages of purple corn lines in Autumn-Winter season 2021 in Gia Lam, Hani. The stage from sowing seeds to spreading pollen.
The stage from sowing seeds to spraying beard 4. The difference between tossing pollen spraying beard. The period from sowing to harvesting fresh corn. The period from sowing to physiological maturity.
Some agronomic characteristics of purple corn lines in Autumn- Winter season 2021 in Gia Lam, Han0i. Ratio of ear height/plant height. cesses eee ceeeeeeeeeneens 49 4.-- - «+ xà SH HH HT TH rep 50 4. Pests and diseases of purple waxy inbred lines in Winter season 2021 at EU.
52 Rhizoctonia solani Kuhn. --- -- 5- <5 SE x3 TH HT HH HT rệt 4. Rhizoctonia solani Kuhn 4. POUT the TOOES .- 1+4 HT HH TH HT TT rệt 4.
Yield and yield components of purple waxy corn inbred line in Winter season 2021 at HanOi.-- - --- - 5: S13 1T TH HH HH TH HH 4. -- -- -- 1 1S 3111111 1111110 11118111 HH HH Hy 59 1H 4. Number of kernel rows per ear (RE). Number of kernels per row (KR) 4.- 2à He, ch so si 0à, yð2.
Quality parameters of of purple waxy corn inbred line in Winter season 2021 at Hanod occ cece ố ố. Correlation between agronomic traits of purple waxy corn lines in Spring 2021 crop in Gia Lam, HanOl. -- - + 5 tt E 9E v3 121v vn ghen 64 4. Selection of elite purple waxy corn inbred lines based in phenotypes.
Genetic diversity of purple corn lines based on SSR marker. 68 PART V: CONCLUSIONS AND RECOMMENDATIONS.---- - + + +2 t3 Ex TT nT KH THHnHHnHHnHrệt 73 458) 545060.cẶ-ĂSS nhe, IV LIST OF TABLES Table 3. Plant materials used In this StUỦy. List of 14 directives used Íor SUTV€Y1TE.- 5-5 Sex seeeseke 39 Table 4.
Growth duration of purple waxy corn inbred line in Winter season at HanoI, 22. Morphological traits of purple waxy corn inbred lines. Pest and disease tolerance of purple waxy corn inbred line in Winter season 202l at HanOI. - -- 5-52 + 1 1E về 1 TT TT HT TH HT HT HH kiệt 52 Table 4.
Yield and yield components of purple waxy corn inbred line in Winter season 2021 at HanOI.-- 5 +5: 2323 3k SH HH Tnhh 55 Table 4. Quality of purple waxy corn inbred line in Winter season 2021 at EU. Genetic similarity matrix LIST OF FIGURES Figure 2. Dried waxy corn containing different types of seeds.
Carbohydrate compositions (mg g"! fresh weight) of six waxy corn 10 Figure 2. Schematic illustration of the WX gene. Kernels color of waxy corn genotypes at full maturity. Diversity 1n COÏOT€d COTT1.
- 5-52 kE*E*k£vHngrvrkt 18 Figure 4. Correlation heatmap of phenotypic traits measurement on purple WaXY COMM inbred LMS TT. Selection of elite purple waxy corn based on phenotypic traits with 25% selection intensity4. Genetic diversity of purple corn lines based on nai Tố.
Cluster dendrogram presented genetic diversity of purple waxy corn based on phenotypic {T1{S. +6 1+4 kg Hành 68 Figure 4. Gel electrometric of 14 SSR markers on 14 elite purple waxy corn inbred lines with check inbred line INÑTÌI I Ì. UPGMA dendrogram based on the SSR markers of 14 elite purple waxy corn inbred lines with check variety INT'Í [1 .---- << +<s++s++xs+e+ 72 Abbreviate al ANOVA CV LAI LSD MAS MGIDI QCVN RCBD Se Sul Sh2 UDP-Glucose VCU LIST OF ACRONYMS Description Anthocyanin biosynthetic gene, anthocyanins-1 Analysis of Variance Coefficient of Variation Leaf Area Index Least Significant Different Marker Assist Selection Multitrait Genotype-Ideotype Distance Index Vietnamese Standards Randomized Complete Block Design Sugary enhanced Sugary 1 Shrunken 2 Uridine diphosphate glucose Value of Cultivation and Use ABSTRACT Limited labor, cost, and time regarding hybrid formation and yield trials encourages corn breeders to identify potential Fl hybrids without crossing all possible combinations by line screening based SSR-genetic distance.
Therefore, this study aimed to determine combining ability and heterosis of parental lines included 55 purple waxy corn inbred lines, denoted from DOI to D55, with check is purple waxy corn inbred line NT111 , to save costs only 14 promising lines + 1 control line, NT111, were selected to analyze genetic diversity based on SSR molecular markers.(parental line of hybrid purple waxy corn VNUA141) developed by the Institute of Crop Research and Development, Vietnam National University of Agriculture. Genetic diversity analysis based on 20 phenotypic traits showed that purple corn lines were divided into 4 main groups that exhibited relatively high levels of genetic diversity. Based on SSR markers , with the coefficient 0.8, those purple waxy corn inbred lines were divided into 4 groups. There were was a poor correlation between phenotypic diversity and genetic divserity of those studied purple waxy corn inbred lines.
Research gap Purple waxy corn (Zea mays L.) is edible with high anthocyanin content (Harakotr et al. Pigments may possess valuable potentials in health product industries as they also contain the bioactive compounds (Limsitthichaikoon et al. There are number of special cultivars that contain colored pigments and give rise to numerous varieties of black and purple corn. The dark purple color of corn is caused by high content of anthocyanins located in the pericarp layers and cob.
Anthocyanin pigment was found in all parts of purple corn, but it was found at particularly high concentration in the husk and cob (Li et al. Kernel pigments like anthocyanins and carotenoids have numerous nutritional functions in animals and human beings. Increasing the levels of these compositional traits and pigments in kernels should increase the nutritional quality of corn (Si Hwan Ryu, 2010). Purple waxy corn cob is one of the interesting agricultural wastes for health products because of aqueous extracts from those were shown to have potential valuable contents for health use (Limsitthichaikoon et al.
Anthocyanin pigment was found in all parts of purple corn, but it was found at a particularly high concentration in the husk and cob (Yousuf et al. High yield is still a primary goal of most plant breeding programs Ferh.5 Pest resistance, stalk strength, uniformity, kernel quality and early maturity are also important in corn and waxy corn breeding programs. Currently, most corn varieties grown commercially are hybrids. However, some purple waxy corm varieties in Asian counties and Vietnam among them are open-pollinated varieties (OPVs) and imported from other countries.
Therefore, improved OPVs and selection of national and exotic germplasm are important for purple waxy hybrid development. Currently, very few systematic studies of the genetic diversity and genetic relationships among purple waxy corn inbred lines have been conducted to date. Since these accessions and inbred lines have not been or are rarely utilized in breeding programs. Genetic characterization is needed to ensure the long-term success of purple waxy corn breeding programs.
Therefore, this study was conducted to investigate the genetic diversity and genetic relationships of purple waxy corn inbred lines by phenotype and SSR markers to for hybrids purple waxy corn breeding program. Objective and requirements 1. Objective Assessment genetic diversity of the purple waxy inbred line revealed by phenotype and SSR markers at Gia Lam, Ha Noi in autumn-winter season 2021, to identify potential inbred lines for development new hybrid purple waxy corn varieties. Requirements - Evaluation of the growth and development stages and some agro- biological characteristics, resistance to pests and diseases, and broken beans of waxy corn lines in the Autumn-Winter season 2021 in Gia Lam, Hanoi.
- Evaluation of yield and yield components of waxy corn lines in Autumn- Winter season 2021 in Gia Lam, Hanoi. - Evaluation of quality criteria of pure waxy corn lines in Autumn-Winter season 2021 in Gia Lam, Hanoi. - Analysis of genetic diversity of pure waxy corn lines Autumn-Winter season 2021 in Gia Lam, Hanoi based on phenotype. - Analysis of genetic diversity of pure waxy corn lines in Autumn-Winter season 2021 in Gia Lam, Hanoi based on SSR markers.
History original of the purple waxy corn In the early 1900s, the waxy corn trait, which is mediated by the WX gene, was found in China. According to James L.