中国科学院海洋研究所机构知识库

植物根结线虫是一类危害严重的农业病原，每年给农业生产造成巨大损失，严重危害了粮食安全性，目前主流的的防治手段多为使用化学农药进行防控，但常用的化学杀线剂多存在高毒、高残留等问题，且随着化学杀线剂的大量使用线虫的抗药性明显增强。因此，开发绿色、高效的杀线制剂已是当务之急。

5-羟甲基糠醛（HMF）是一种具有广泛应用前景的生物基平台化合物，但此前的研究中多使用可食用性生物质如果糖，葡萄糖等为原料生产HMF，面临着与人争粮的问题，而海洋中丰富的甲壳废物资源，是制备HMF的潜在理想原料，具有重要的研究意义和价值。前期有少量文献报道了HMF在离体条件下具有较好的杀线活性，但缺乏进一步的活体防效验证。而且由于HMF对光、热、酸等敏感，其储存、使用存在诸多不便，因而极大的限制了其进一步开发与应用。而膨润土是一类具有多种优异性能的功能型层状粘土，具有丰富的矿藏量，价格低廉，具有较强的吸附性能，在农业上广泛应用于各种农药和肥料的载体和助剂。通过对HMF不稳定性机理的分析，本论文创新性的使用膨润土作为HMF的载体制备了一种HMF颗粒剂，并对其进行了系统的结构表征和理化性质研究，评价了其稳定性和对南方根结线虫的杀线活性，本文完成的工作简述如下：

1.设计并发展了一种以氨基葡萄糖为原料高效催化转化制备HMF的方法和体系，HMF产率可达42.13%，接近已报道的以氨基葡萄糖为原料合成HMF的最高产率；经高效液相法验证，未经处理的反应产物中，HMF的纯度＞96%，表明该反应体系具有较好的选择性。

2.以膨润土为载体负载HMF，制备了一种复合载药体，即HMF颗粒剂，在常温下储存的三个月内有效成分的含量未发生减少，极大的改善了HMF在常温下易分解变质的缺陷，为HMF开拓了更广阔的应用前景。

3.对制备的HMF颗粒剂的多项结构表征进行了测试，通过对XRD、热重测试、接触角测试和电镜图像的分析，提出了膨润土吸附HMF赋予其稳定性的猜想。

4.在盆栽实验中验证了HMF颗粒剂对南方根结线虫的防治活性，HMF颗粒剂对南方根结线虫具有良好的防效，为控制根结线虫病害提供了新策略。

Plant root-knot nematodes are a serious agricultural pathogen that cause immense losses to agricultural production annually, posing a significant threat to food security. Currently, the main method of control involves the use of chemical pesticides. However, commonly used nematicides often have issues such as high toxicity and residue, and the overuse has led to an increase in nematode resistance to these chemicals. Therefore, developing green and efficient nematicides is an urgent task.

5-Hydroxymethylfurfural (HMF) is a bio-based platform compound with wide-ranging applications. Previous studies have primarily used edible biomass such as fructose and glucose to produce HMF, which competes with food supply. However, abundant crustacean waste resources in the oceans are potentially ideal raw materials for the preparation of HMF, holding significant research significance and value. There are a few reports of HMF's nematicidal activity under in vitro conditions, but further in vivo efficacy validation is lacking. Moreover, due to HMF's sensitivity to light, heat, and acid, its storage and use are inconvenient, which significantly limits its further development and application. Bentonite is a type of functional layered clay with many excellent properties, abundant deposits, and low cost. It has strong adsorption properties and is widely used in agriculture as a carrier and adjuvant for various pesticides and fertilizers. By analyzing the instability mechanism of HMF, this thesis innovatively uses bentonite as a carrier for HMF, preparing a form of HMF granules. It includes structural characterization and physicochemical properties research of these granules, studying their stability, and investigating their nematicidal activity against Meloidogyne incongnita. The work completed in this paper is briefly summarized as follows:

1. A method and system were designed and developed for the efficient catalytic conversion of glucosamine into HMF, achieving a yield of 42.13%, close to the highest yield reported for HMF synthesis using glucosamine. Verified by HPLC test, the purity of HMF in the untreated reaction product was over 96%, indicating good selectivity of the reaction system.

2. A composite pesticide carrier, namely HMF granules, was prepared using bentonite as the carrier. The active ingredient content did not decrease within three months of storage at room temperature, significantly improving the easy decomposition and deterioration of HMF at room temperature and expanding its application prospects.

3. Various structural characterizations of the prepared HMF granules were tested. Through analysis of XRD, thermogravimetric tests, contact angle tests, and electron microscope images, a hypothesis was proposed on how bentonite adsorption imparts stability to HMF.

4. The nematicidal activity of HMF granules against the Meloidogyne incongnita was validated in pot experiments. The HMF granules showed good efficacy, providing a new strategy for controlling root-knot nematode diseases.

目  录

第1章 绪论 1

1.1 线虫 1

1.1.1 线虫概述 1

1.1.2 根结线虫的形态和生活史 1

1.1.3 根结线虫的危害 2

1.1.4 根结线虫的防治难点 3

1.1.5 根结线虫防治措施 3

1.2 甲壳资源防治根结线虫的研究 6

1.2.1 甲壳源大分子化合物 7

1.2.2 甲壳源小分子化合物 7

1.3 膨润土 11

1.3.1 膨润土和粘土矿物概述 11

1.3.2 膨润土的应用 11

1.3.3 膨润土作为农药载体的优势 13

1.4 选题思路 15

1.5 选题的目的和意义 16

1.6 研究技术路线与内容 17

第2章 HMF的制备与载药体系构建 19

2.1 材料与仪器 19

2.1.1 材料 19

2.1.2 实验仪器 19

2.2 实验方法 19

2.2.1 壳聚糖及氨基葡萄糖催化转化制备HMF研究 19

2.2.2 HMF的HPLC检测 20

2.2.3 HMF颗粒剂的制备 20

2.2.4 HMF颗粒剂含量检测 20

2.3 结果与分析 21

2.3.1 HMF的制备的产率与选择性 21

2.3.2 HMF颗粒剂的制备 24

2.4 讨论 25

第3章 HMF颗粒剂的稳定性的研究与结构表征 29

3.1 材料与仪器 29

3.1.1 材料 29

3.1.2 实验仪器 29

3.2 实验方法 29

3.2.1 HMF颗粒剂稳定的研究 29

3.2.2 HMF颗粒剂结构表征 29

3.2.3 HMF分子尺寸的计算 30

3.3 结果与分析 30

3.3.1 HMF颗粒剂稳定性研究的结果 30

3.3.2 HMF颗粒剂结构表征的结构 32

3.4 讨论 37

第4章 HMF颗粒剂对根结线虫的盆栽活性试验及植物安全性初步研究 41

4.1 材料与仪器 41

4.1.1 材料 41

4.1.2 实验仪器 41

4.2 实验方法 41

4.2.1 根结线虫2龄幼虫的孵化和收集 41

4.2.2 黄瓜植株的培育 41

4.2.3 盆栽处理与移栽 42

4.2.4 发病率统计与防效计算 42

4.2.5 植物生长指标的测量 42

4.3 结果与分析 42

4.3.1 HMF颗粒剂在盆栽试验中对根结线虫的防治效果 43

4.3.2 HMF颗粒剂对植物生长指标的影响 45

4.4 讨论 50

第5章 结论与创新点 53

5.1 本文结论 53

5.2 创新点 53

参考文献 55

致  谢 65

作者简历及攻读学位期间发表的学术论文与其他相关学术成果 67