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Allelopathic Effects of Prunus Cerasoides Buch.-Ham Ex. D

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Allelopathic Effects of Prunus Cerasoides Buch.-Ham Ex. D Allelopathic Effects of Prunus cerasoides Buch.-Ham ex. D. Don Leaves on Common Weeds in Forest Restoration Sites Punnat Changsalak Bachelor of Science Biology Department of Biology, Faculty of Science Chiang Mai University i Acknowledgement This research could not have been finished without the help of my Advisor, FORRU- CMU staff and friends. I would like to thank you all of my success in this step. First, I would like to thank my Advisor, Dr. Stephen Elliott, for his invaluable help on this research, especially in writing period. Similarly, I am also grateful to other advisor, including Assistant Professor Dr. Sutthathorn Chairuangsri and Assistant Dr. Pimonrat Tiansawat. Thanks due to The Development and Promotion of Science and Technology Talented Project (DSTP), for financial support during studying in bachelor degree at Chiang Mai University. I wish to express my appreciativeness for The Forest Restoration Research Unit (FORRU- CMU) for facilities support during research. I thank to all FORRU-CMU staff who helped with experiments in the nursery as well. In addition, Thanks are due to Bhanuwat Foojakham, Kotchamon Chamhar, Wittawat Insian and Sarana Chansuebsri, who encouraged and helped me in material collection from the field. Furthermore, I would like to acknowledge the Department of Biology, Faculty of Science, Chiang Mai University which gave me a lot of new experiences and beneficial knowledge for the further education. ii Research Title Allelopathic effects of Prunus cerasaides Buch.-Ham ex. D. Don leaves on common weeds in forest restoration site Author Mister Punnat Chnagsalak B.S. Biology Examining Committee Lecturer Dr. Stephen Elliott Chairperson Assist. prof. Dr. Sutthathorn Chairuangsri Member Assist. Dr. Pimonrat Tiansawat Member Abstract Weeding by hand tools is the most expensive part of forest restoration projects, so herbicides are an attractive alternative weeding technique. However, synthetic herbicides may have negative impacts on the environment and human health. Prunus cerasoides Buch.-Ham. Ex D.Don produces allelochemicals, which may enable the tree to compete with herbaceous weeds such as Chromoleana odorata (L.) R.M.King & H.Rob. and Bidens pilosa L.. Consequently, it may be possible to develop more environment-friendly “bio”-herbicides from P. cerasoides allelochemicals. This project investigated the inhibitory potential of aqueous leaf extract (ALE) from P. cerasoides leaves on seed germination and seedling growth of C. odorata and B. pilosa - two abundant weeds of forest restoration sites. ALE at various concentrations (0.75-5.00 wt%) was applied to weed seeds to test the intensity and duration of its inhibitory effect on germination. ALE at 0.75 and 1.25 wt% significantly inhibited germination of C. odorata and B. pilosa, respectively (P≤0.05), with the degree of inhibition increasing with increasing concentration. It also delayed germination for a few days (1-4 days). ALE had no significant substantial inhibitory effect on seedling survival and biomass per plant irrespective of development stage. Consequently, P. cerasoides ALE should be further investigated as a pre-emergent herbicide. It is unlikely to be useful as a general weed killer on forest restoration sites. iii หัวข้อปัญหาพิเศษ ผลจากอัลลีโลพาธีของใบ Prunus cerasoides Buch.-Ham ex. D. Don ต่อวัชพืช ที่พบได้บ่อยภายในแปลงฟื้นฟู ชื่อผู้แต่ง นายปุณณัตถ์ ช่างสลัก วิทยาศาสตรบัณฑิต สาขาวิชาชีววิทยา คณะกรรมการสอบปัญหาพิเศษ อ. ดร. สตีเฟน เอลเลียต ประธานกรรมการ ผศ. ดร. สุทธาธร ไชยเรืองศรี กรรมการ ผศ. ดร. พิมลรัตน์ เทียนสวัสดิ์ กรรมการ บทคัดย่อ การกำจัดวัชพืชโดยการตัดด้วยเครื่องมือกำจัดวัชพืชแบบทั่วไปเป็นค่าใช้จ่ายที่สำคัญในโครงการฟื้นฟู สารกำจัดวัชพืชจึงเป็นทางเลือกที่น่าสนใจ แต่งานวิจัยพบว่าสารกำจัดวัชพืชอาจส่งผลกระทบต่อสิ่งแวดล้อม และสุขภาพของมนุษย์ Prunus cerasoides Buch.-Ham. Ex D.Don มีกลไกอัลลีโลพาธีในการต่อสู้กับวัชพืช พื้นล่างที่มีอยู่ในตามธรรมชาติ หากนำความสามารถนี้ของ P. cerasoides มาใช้พัฒนาเป็นสารกำจัดวัชพืชที่ เป็นมิตรต่อสิ่งแวดล้อม อาจเป็นประโยชน์แก่โครงการการฟื้นฟูป่าในลำดับต่อไป ปัญหาพิเศษนี้จึงจัดทำขึ้น เพื่อทดสอบความสามารถของสารสกัดหยาบจากใบ (ALE) ของ P. cerasaides ต่อการงอกของเมล็ดและการ เจริญเติบโตของต้นกล้าวัชพืชที่พบได้บ่อยในแปลงฟื้นฟู ได้แก่ C. odorata และ B. pilosa โดยใช้ ALE ระดับ ความเข้มข้นตั้งแต่0.75-5.00 wt% ในการทดสอบการยับยั้งการงอกเมล็ดวัชพืช ทำให้ทราบถึงระดับความ เข้มข้นที่มีความสามารถยับยั้งเมล็ด C. odorata และ B. pilosa ได้ดีที่สุด คือ 0.75 และ 1.25 wt% ตามลำดับ และผลจาก ALE ยังสามารถชะลอการงอกของเมล็ดทั้ง 2 ชนิดได้ (ประมาณ 1-4 วัน) จากนั้นใช้ ระดับความเข้มข้นที่ได้ทดสอบกับต้นกล้าอายุ 2 เดือนของวัชพืช ทำให้ทราบว่า ALE ไม่มีผลต่ออัตราการตาย iv และน้ำหนักแห้งของต้นกล้าทั้ง 2 ชนิด จึงมีการเพิ่มระดับความเข้มขึ้นเป็น 4 เท่าจากเดิม (ซึ่งระดับความ เข้มข้นที่ใช้กับต้นกล้า C. odorata และ B. piloas คือ 3.00 และ 5.00 wt% ตามลำดับ) เพื่อทดสอบกับต้น กล้าวัชพืชที่มีระยะการพัฒนาแตกต่างกัน ซึ่งที่ความเข้มข้นดังกล่าว มีผลทำให้เกิดการเปลี่ยนแปลงอัตราการ ตายและน้ำหนักแห้งในต้นกล้าบางระยะของ C. odorata เท่านั้น ในขณะที่ไม่พบการเปลี่ยนแปลงใด ๆ ในต้น กล้า B. pilosa ทุกๆระยะ จากการศึกษาครั้งนี้ ทำให้ทราบว่า ALE มีความสามารถในการยับยั้งการงอกของ เมล็ดวัชพืชแต่ไม่มีผลต่อต้นกล้าวัชพืชจึงยังไม่เหมาะสมสำหรับการใช้ควบคุมวัชพืชในแปลงฟื้นฟู v Table of Contents Page Acknowledgement i Abstract in English ii Abstract in Thai iii Table of Content v List of Tables vii List of Figures viii Chapter 1 Introduction 1 1.1 Introduction 1 1.2 Objective 2 Chapter 2 Literature review 3 2.1 Chemical herbicides 3 2.2 Allelopathy and allelochemicals 4 2.3 Chemicals isolated from the P. cerasoides 5 2.4 Forest restoration 6 Chapter 3 Material and method 8 3.1 Preparation of Aqueous leaf extract (ALE) 8 3.2 Application of aqueous leaf extract (ALE) 8 3.3 Application of acidic treatment 9 3.4 Selected weed collection 9 3.5 Preparation of seedlings in nursery. 9 3.6 Experimental Design 10 3.7 Dry weight measurement 11 vi Table of Contents (Continued) Page 3.8 Seedling height measurement 11 3.9 Data analysis 12 Chapter 4 Results 13 4.1 Seed germination 13 4.2 Seedling growth 16 Chapter 5 Discussion 21 4.1 Seed germination 21 4.2 Seedling growth 22 Chapter 6 Conclusion 24 References 25 Appendices 30 Appendix A Pictures about the experiment 31 Appendix B Seed germination data 35 Appendix C Seedling growth data 38 Curriculum Vitae 45 vii List of Tables Tables Page 1 Amount of water and powder in different concentration treatment 8 2 Percent of seed germination of C. odorata after 9 days 38 3 Percent of seed germination of B. pilosa after 9 days 38 4 Cumulative germination of C. odorata seeds over 14 days 39 5 Percent of seed germination of C. odorata after 14 days 39 6 Cumulative germination of B. pilosa seeds over 14 days 40 7 Percent of seed germination of B. pilosa after 14 days 40 8 Percent mortality of C. odorata seedlings after 31 days 41 9 Percent mortality of B. pilosa seedlings after 31 days 41 10 Biomass per plant of C. odorata seedlings after 31 days 41 11 Biomass per plant of B. pilosa seedlings after 31 days 41 12 Percent mortality of C. odorata in each development stage seedlings 42 after 15 days 13 Percent mortality of B. pilosa in each development stage seedlings 42 after 15 days 14 Biomass per plant of C. odorata in each development stage seedlings 43 after 15 days 15 Biomass per plant of B. pilosa in each development stage seedlings 43 after 15 days 16 Seedling height of C. odorata in each development stage after 15 days 44 17 Seedling height of B. pilosa in each development stage after 15 days 44 viii List of Figures Figure Page 1 Dried leaves, grinding the leaves into powder and sieving to make fine powder 8 2 Seedlings after transplanted into germination trials 9 3 All Experimental design diagrams 11 4 Germination percentage of C. odorata seeds after 9 days 13 5 Germination percentage of B. pilosa seeds after 9 days 14 6 Cumulative germination of C. odorata seeds over 14 days 15 7 Cumulative germination of B. pilosa seeds over 14 days 15 8 Mortality of C. odorata and B. pilosa seedlings over 31 days 16 9 Biomass per plant of C. odorata and B. pilosa seedlings after 31 days 17 10 Mortality of C. odorata seedlings and B. pilosa seedlings over 15 days 18 11 Biomass of C. odorata seedlings after 15 days 19 12 Biomass per plant of B. pilosa seedlings after 15 days 19 13 Seedling height of C. odorata seedlings after 15 days 20 14 Seedling height of B. pilosa seedlings after 15 days 20 15 C. odorata and B. pilosa seedlings 34 16 P. cerasoides flower, bark and leaves 34 17 P. cerasoides dried leaves 35 18 Germinated C. odorata and B. pilosa seeds. 35 19 P. cerasoides ALE powder, before mixing with water and after mixing of water 36 20 B. pilosa seeds treated with P. cerasoides ALE, compared with control and 36 acidic treatment 21 Experimental Design of B. pilosa 36 ix List of Figures (Continue) Figure Page 22 Comparison of C. odorata seedlings from control, acidic and ALE treatment, 37 at the end of Experiment III (after 31 days). 23 Comparison of B. pilosa seedlings from control, acidic and ALE treatment, 37 at the end of Experiment III (after 31 days). 1 Chapter 1 Introduction 1.1 Introduction Weed growth is one of the most serious factors that prevent the success of forest restoration projects, particularly invasive exotic weed species. Weeds inhibit growth and survival of both naturally regenerating and planted trees, due to competition for nutrients, water and light (Harper 1982). Some of the most ubiquitous weeds that impede forest restoration in northern Thailand are Chromoleana odorata (L.) R.M.King & H.Rob. and Bidens pilosa L. (both
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