植物面對重金屬逆境的反應，含括複雜的機制及接受重金屬訊息而活化的訊息傳遞途徑、生合成逆境相關的蛋白質及訊息傳遞分子，以及專一的重金屬反應相關基因轉錄活化以應對逆境。六價鉻［Cr（VI）］對於一般植物而言是一種非生長必需的金屬，當濃度過高時則會造成毒害。然而目前對於鉻影響細胞功能及基因表現的訊息傳遞途徑及分子機制所知有限。在本篇研究中，我們證實鉻會誘導細胞內的過氧化物（ROS）產生以及鈣離子（Ca2+）累積。透過微陣列晶片分析，我們研究水稻根部中的全面性基因轉錄變化。由鉻誘導水稻根部內基因表現特性顯示離層酸、乙稀及茉莉酸訊息傳遞與生合成相關基因有表現量增加的現象，而吉貝素則有失去活性的情形。在鉻逆境中，訊息分子例如蛋白質磷酸化酵素（含未知功能區域26）、類受質胞質磷酸化酵素、類LRR10磷酸化酵素種類2及蛋白質去磷酸化酵素2C，以及轉錄分子例如WRKY及apetala2/乙稀反應因子，其基因表現量特別明顯。特別的是，一些參與囊泡運輸的基因也是受鉻刺激而大量表現。當預先以囊泡運輸抑制劑 （brefeldin A）處理水稻根部時，會有效的減少鉻所誘導的ROS產生。
為了暸解囊泡運輸在重金屬逆境中的角色，我們挑選參與囊泡運輸的基因以進行功能確認，NbExo70啟動子/GUS報導蛋白活性確實會受六價鉻或二價銅逆境所誘導。藉由共軛焦顯微鏡，我們知道NbExo70蛋白在細胞中的位置於細胞膜及細胞質。利用病毒誘導基因靜默（VIGS）抑制參與囊泡運輸的基因NbExo70，顯示重金屬誘導的ROS產生，需要囊泡運輸的參與。此外，當大量表現缺乏區域D（其作用為影響NbExo70膜交互作用）的NbExo70時，一樣會顯著地降低重金屬與鹽誘導植物產生ROS。特別的是，NbExo70的靜默與區域移除突變會增加植物的重金屬與鹽逆境容忍度。這些結果提供我們對於植物面對環境逆境時，其訊息傳遞、基因調控及囊泡運輸所扮演的角色更多暸解。

The plant response to heavy metal stress involves a complicated mechanism and signal transduction network that are activated by sensing the heavy metal, and are characterized by the synthesis of stress-related proteins and signaling molecules, and the finally the transcriptional activation of specific metal-response genes to counteract the stress. Hexavalent chromium [Cr (VI)] is a non-essential metal for normal plants and is toxic to plants at high concentrations. However, little is known about the signaling pathways and molecular mechanisms of its action on cell function and gene expression. In this study, we demonstrated that Cr (VI) induced intracellular reactive oxygen species (ROS) production and Ca2+ accumulation. We investigated global transcriptional changes in rice roots by microarray analysis. Gene expression profiling of Cr (VI)-treated rice roots revealed upregulated signaling and biosynthesis of abscisic acid, ethylene, and jasmonic acid and inactivation of gibberellic acid. Genes encoding signaling components such as the protein kinases domain of unknown function 26, receptor-like cytoplasmic kinase, LRK10-like kinase type 2, and protein phosphatase 2C as well as transcription factors WRKY and apetala2/ethylene response factor were predominant during Cr (VI) stress. Specially, several genes involved in vesicle trafficking were also upregulated. Pre-treating rice roots with a vesicle trafficking inhibitor, brefeldin A, effectively reduced Cr (VI)-induced ROS production.
To find out the role of vesicle trafficking in heavy metal stress response, genes involved in vesicle trafficking were subjected to functional characterization. NbExo70 promoter/GUS reporter activity was strongly induced by Cr (VI) or Cu (II) stress. Confocal microscopy indicated that NbExo70 protein is located both at the plasma membrane and within the cytoplasm. Suppression of the vesicle trafficking-related gene, NbExo70, by virus-induced gene silencing (VIGS) strategies revealed that vesicle trafficking is required for mediation of heavy metal-induced ROS production. Furthermore, overexpression of dominant negative form of NbExo70 domain D, which may result in disrupting the NbExo70 membrane interaction, also showed a significantly decrease in heavy metal and salt induced-ROS production in plants. Specially, the silencing and dominant negative mutation of NbExo70 exhibited an increased tolerance to heavy metal and salt stress in plants. These results provide insights into understanding the molecular mechanisms in signaling pathways, transcriptional regulation and the role of vesicle trafficking in response to environmental stress in plants.

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