玉米根边缘细胞exDNA和胞外蛋白对土荆芥化感胁迫的缓解效应

doi:10.3864/j.issn.0578-1752.2015.10.009

摘要/Abstract

摘要： 【目的】根边缘细胞（root border cells，RBCs）是从根冠上脱落下来的单个细胞或小细胞团。由于根边缘细胞的黏胶层与哺乳动物中性粒细胞胞外诱捕网（NETs）具有类似的防御功能，而被称为根边缘细胞胞外诱捕网（BETs）。exDNA和胞外蛋白是根边缘细胞黏胶层的组成成分，在根边缘细胞抵御生物和非生物胁迫中起着重要作用。化感作用是外来植物成功入侵的机制之一，探讨exDNA和胞外蛋白在根边缘细胞黏胶层抵御入侵植物土荆芥化感胁迫中的作用，有助于深入研究植物根边缘细胞抵御化感胁迫的机制。【方法】以在土荆芥入侵地广泛种植的农作物玉米（雅玉26^#）为材料，采用纯琼脂培养基悬空气培养法，研究供试玉米品种（雅玉26^#）根边缘细胞的发育特性，以及在土荆芥挥发油及其两种主要成分对伞花素和α-萜品烯作用下，玉米根边缘细胞活性和黏胶层相对面积的变化规律，并分析在根边缘细胞黏胶层exDNA或胞外蛋白被DNA酶或蛋白酶降解的情况下，土荆芥挥发油及其主要成分对伞花素以及α-萜品烯对根边缘细胞活性和黏胶层相对面积的影响。【结果】供试玉米根边缘细胞几乎与根尖同时出现，并随着根的伸长而逐渐增加，当根长达到30 mm时，根边缘细胞数量达到最大值，约6 130个。之后，随着根进一步伸长，根边缘细胞数量略微减少并趋于平衡，而根冠PME活性与根长呈现极显著的负相关（P＜0.01）。根冠果胶甲基酯酶（PME）活性在根刚露白时就处于相对高的水平，之后则随着根伸长PME活性呈现持续下降最终趋于稳定；土荆芥挥发油、对伞花素、α-萜品烯具有细胞毒性，均能导致根边缘细胞活性显著下降（P＜0.05），其中，挥发油的毒性最大，对伞花素、α-萜品烯的毒性次之，对伞花素和α-萜品烯混合物的毒性最小；土荆芥挥发油具有诱导根边缘细胞黏胶层面积增大的效应，并表现出剂量效应，当挥发油剂量达到5 μL时，黏胶层相对面积与对照差异显著（P＜0.05）。与对照相比，在对伞花素和α-萜品烯的作用下玉米根边缘细胞黏胶层面积变化不显著；当exDNA或胞外蛋白被DNA酶或蛋白酶降解后，根边缘细胞黏胶层相对面积缩小，细胞活性降低，其中，挥发油处理组根边缘细胞活性均在10%以下，对伞花素处理组、α-萜品烯处理组、对伞花素和α-萜品烯混合物处理组根边缘细胞活性虽然有所降低，但仍保持在80%左右。【结论】土荆芥挥发油及其主要成分对伞花素、α-萜品烯具有细胞毒性，可导致玉米根边缘细胞活性降低，根边缘细胞黏胶层中的exDNA和胞外蛋白对土荆芥挥发油、对伞花素、α-萜品烯的细胞毒性具有缓解效应，可在一定程度上缓解土荆芥的化感胁迫。

关键词: 土荆芥, 化感胁迫, exDNA, 胞外蛋白, 玉米根边缘细胞, 缓解效应

Abstract: 【Objective】Root border cells (RBCs) are released from the root cap as individual cells or a group of attached cells. The mucilage of root border cells acts in a manner similar to that of neutrophil extracellular traps (NETs) in defense, thus, known as border cell extracellular traps (BETs). The extracellular DNA (exDNA) and proteins are components of border cell mucilage, which have been considered to play a vital role in protecting root tip from biotic and abiotic stresses. Allelopathy is one of the successful mechanisms of exotic plant for invading. The objective of this study is to reveal the roles of the exDNA and extracellular proteins of root border cell mucilage in resistance to allelochemical stress from an invasive plant, Chenopodium ambrosioides L..【Method】 Experiments were performed with maize (Zea mays L.) ‘Yayu26^#’, a widely grown crop in the introduced habitats of C. ambrosioides under aeroponic culture with agar medium. The developmental characteristics of root border cells in maize were investigated. And the roots were treated with volatile oil from C. ambrosioides and its two main components, α-terpinene and cymene, respectively, then the experiments were divided into two groups. In one group, the root border cell viabilities and relative areas of root border cell mucilage were measured directly. In another group, the exDNA or extracellular proteins of root border cell mucilage were degraded by DNase I or protease, and then the cell viabilities and relative areas of root border cell mucilage of maize were studied.【Result】 The first border cell formation almost synchronized with root tip emergence in maize. The number of border cells increased with the development and elongation of the root. When the root length was about 30 mm, the number of border cells reached the maximum, that was about 6 130, then declined slightly to a stable level. Meanwhile, the activities of pectin methylesterase were correlated with root length negatively (P＜0.01). Once the emerging of root, the activities of pectin methylesterase reached the top, and then declined into a low level. The cytotoxicity from volatile oil, α-terpinene and cymene resulted in significant decrease of the viabilities of root border cells (P＜0.05). The allelopathic effects of α-terpinene and cymene were weaker than that of volatile oil. The allelopathic effects of mixed treatment group were different from the individual groups, which the viabilities of root border cells were higher than any other treatment groups, showed minimal toxicity. Volatile oil from C. ambrosioides had potential to induce the enlargement of mucilage with dose-dependent. At the dose of 5 μL, the relative areas of mucilage enlarged significantly in volatile oil treatment groups (P＜0.05). Compared with the control, α-terpinene-treated and cymene-treated induced no significant change of the relative areas of mucilage. Degradation of exDNA or extracellular proteins with DNase I or protease resulted in loss of root border cells resistance to allelochemical stress, and the viability of root border cells decreased obviously, there was less than 10% viabilities of root border cells remained in volatile oil treatment groups, while there was more than 80% in α-terpinene-treated, cymene-treated and the combined treatment of α-terpinene and cymene. 【Conclusion】Volatile oil from C. ambrosioides and its two main components, α-terpinene and cymene were cytotoxical, both of them inhibited viabilities of root border cells, while exDNA and extracelluar proteins of root border cell mucilage play an important role in resistance to allelochemical stress from C. ambrosioides.

Key words: Chenopodium ambrosioides L., allelochemical stress, exDNA, extracellular protein, root border cells of maize, alleviating effect

胡忠良，王亚男，马丹炜，陈斌，何亚强，周健. 玉米根边缘细胞exDNA和胞外蛋白对土荆芥化感胁迫的缓解效应[J]. 中国农业科学, 2015, 48(10): 1962-1970.

HU Zhong-liang, WANG Ya-nan, MA Dan-wei, CHEN Bin, HE Ya-qiang, ZHOU Jian. The Alleviate Effect of Extracellular DNA and Protein in Maize Root Border Cells on the Allelochemical Stress from Chenopodium ambrosioides L.[J]. Scientia Agricultura Sinica, 2015, 48(10): 1962-1970.

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