NMT是基因功能的活體檢測技術(shù)，已被31位諾貝爾獎得主所在單位，及北大、清華、中科院使用。
期刊：Chemosphere
主題：浮萍超富集Cd的分子機制研究
標題：Comparative transcriptome analysis of duckweed (Landoltia punctata) in response to cadmium provides insights into molecular mechanisms underlying hyperaccumulation
影響因子：4.427
檢測指標：Cd2+流速
檢測樣品：浮萍根（根尖、距離根尖400μm，1000μm）
Cd2+流速流實驗處理方法：
50 μM CdCl2處理30min
Cd2+流速流實驗測試液成份：
0.1 mM KCl，0.1 mM MgCl2、0.05 mM CaCl2、0.3 mM MES、50μM CdCl2，pH5.5
作者：中科院青島生物能源與過程研究所周功克、徐華
英文摘要
Cadmium (Cd) is a detrimental environmental pollutant. Duckweeds have been considered promising candidates for Cd phytoremediation. Although many physiological studies have been conducted, the molecular mechanisms underlying Cd hyperaccumulation in duckweeds are largely unknown.
In this study, clone 6001 of Landoltia punctata, which showed high Cd tolerance, was obtained by large-scale screening of over 200 duckweed clones. Subsequently, its growth, Cd flux, Cd accumulation, and Cd distribution characteristics were investigated. To further explore the global molecular mechanism, a comprehensive transcriptome analysis was performed. For RNA-Seq, samples were treated with 20 μM CdCl2 for 0, 1, 3, and 6 days.
In total, 9,461, 9,847, and 9615 differentially expressed unigenes (DEGs) were discovered between Cd-treated and control (0 day) samples. DEG clustering and enrichment analysis identified several biological processes for coping with Cd stress. Genes involved in DNA repair acted as an early response to Cd, while RNA and protein metabolism would be likely to respond as well.
Furthermore, the carbohydrate metabolic flux tended to be modulated in response to Cd stress, and upregulated genes involved in sulfur and ROS metabolism might cause high Cd tolerance. Vacuolar sequestration most likely played an important role in Cd detoxification in L. punctata 6001. These novel findings provided important clues for molecular assisted screening and breeding of Cd hyperaccumulating cultivars for phytoremediation.
中文摘要（谷歌機翻）
鎘（Cd）是有害的環(huán)境污染物。浮萍被認為是鎘植物修復的有前途的候選人。盡管已進行了許多生理研究，但浮萍中Cd過度富集的分子機制尚不清楚。
在這項研究中，通過大規(guī)模篩選200多個浮萍克隆，獲得了對鎘具有高耐受性的Landoltia punctata克隆6001。隨后，研究了其生長，Cd通量，Cd積累和Cd分布特征。為了進一步探索全局分子機制，進行了全面的轉(zhuǎn)錄組分析。對于RNA-Seq，樣品用20μMCdCl2處理0、1、3和6天。
在經(jīng)鎘處理的樣品和對照（0天）樣品之間總共發(fā)現(xiàn)了9,461、9,847和9615個差異表達的單基因（DEG）。 DEG聚類和富集分析確定了應對Cd脅迫的幾種生物學過程。參與DNA修復的基因是對Cd的早期反應，而RNA和蛋白質(zhì)代謝也可能也有響應。
此外，碳水化合物的代謝通量傾向于響應Cd脅迫而被調(diào)節(jié)，而參與硫和ROS代謝的基因上調(diào)可能導致對Cd的高耐受性。液泡螯合最有可能在點球藻6001的Cd解毒中發(fā)揮重要作用。這些新發(fā)現(xiàn)為分子輔助篩選和育種Cd超富集植物進行植物修復提供了重要線索。
結(jié)果表明：葉片（F）的Cd2+吸收速率高于根冠（R0和R1），低于根伸長區(qū)（R2）。葉片中，從節(jié)點（F2）到頂點（F0）的Cd2+吸收速率逐漸下降。根和葉片的Cd2+吸收速率顯著較低,從而更容易適應Cd脅迫。
文章鏈接：
https://www.sciencedirect.com/science/article/pii/S0045653517315680?via%3Dihub