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OsCIPK2過(guò)表達(dá)促進(jìn)低氮條件下水稻NO3-?吸收
NMT是基因功能的活體檢測(cè)技術(shù)，已被103位諾貝爾獎(jiǎng)得主所在單位，及北大、清華、中科院使用。
期刊：International Journal of Molecular Sciences
主題：OsCIPK2過(guò)表達(dá)促進(jìn)低氮條件下水稻NO3-吸收
標(biāo)題：Exploring the Potential of Overexpressed OsCIPK2 Rice as a Nitrogen Utilization E cient Crop and Analysis of Its Associated Rhizo-Compartmental Microbial Communities
影響因子：4.183
檢測(cè)指標(biāo)：Ca2+、NH4+、NO3-流速
檢測(cè)樣品：水稻根尖
Ca2+、NH4+、NO3-流實(shí)驗(yàn)處理方法：
幼苗長(zhǎng)至三葉期，分別在正常氮（1.44 mM NH4NO3）和低氮（0.24 mM NH4NO3）處理
Ca2+、NH4+、NO3-流實(shí)驗(yàn)測(cè)試液成份：
Ca2+: 0.1 mmol/L KCl, 0.1 mmol/L CaCl2, 0.1 mmol/L MgCl2, 0.5 mol/L NaCl, 0.3 mmol/L MES, 0.2 mmol/L Na2SO4, pH 6.0;
NO3-: 0.1 mmol/L NH4NO3, 0.1 mmol/L NH4NO3, 0.1 mmol/L KCl, 0.1 mmol/L KCl, 0.1 mmol/L KCl2, 0.3 mmol/CaCl2, 0.3 mmol/L MgCl2, 0.3 mmol；
NH4+: 0.1 mmol/L NH4NO3, 0.1 mmol/L CaCl2, 0.3 mmol/L MES, pH 6.0
作者：福建農(nóng)林大學(xué)林文雄
英文摘要
Nitrogen (N) is one of the indispensable factors in rice growth and development. China holds a premier position in the production of rice and at the same time also faces higher N fertilizer costs along with serious damage to the environment. A better solution is much needed to address these issues, without disrupting the production of rice as an important cereal, while minimizing all the deleterious effects on the environment.
Two isogenic lines Kitaake (WT) and its genetically modified line CIPK2 (RC), overexpressing the gene for Calcineurin B-like interacting protein kinase 2 (OsCIPK2) with better nitrogen use efficiency (NUE), were compared for their growth and development under low versus normal levels of N. NUE is a complex trait mainly related to a plant’s efficiency in extraction, assimilation, and recycling of N from soil. The microbial population was analyzed using high-throughput Illumina Miseq 16S rRNA sequencing and found that RC with CIPK2, specifically expressed in rice root, not only performed better without nitrogen fertilizer (LN) but also increased the diversity of bacterial communities in rice rhizosphere compartments (rhizosphere, rhizoplane, and endosphere).
The relative abundance of beneficial bacteria phyla increased, which are known to promote the circulation and transformation of N in rhizosphere soil. To further explore the potential of RC regarding better performance under LN, the ion fluxes in root apical were detected by non-invasive micro-test technique (NMT). We found that RC can absorb more Ca2+ and NO3− under LN as compared to WT. Finally, compared to WT, RC plants exhibited better growth of root and shoot, and increased yield and N uptake under LN, whereas there was no significant difference in the growth of two rice lines under normal nitrogen (NN) treatment.
We are able to get preliminary results, dealing with the OsCIPK2 overexpressed rice line, by studying the rice molecular, physiological, and chemical parameters related to NUE. The results laid the foundation for further research on N absorption and utilization in rice from the soil and the interaction with microbial communities.
中文摘要（谷歌機(jī)翻）
氮（N）是水稻生長(zhǎng)發(fā)育*的因素之一。中國(guó)在大米生產(chǎn)中處于地位，同時(shí)也面臨著更高的氮肥成本以及對(duì)環(huán)境的嚴(yán)重破壞。迫切需要一種更好的解決方案來(lái)解決這些問(wèn)題，同時(shí)又不中斷作為重要谷物的大米的生產(chǎn)，同時(shí)將對(duì)環(huán)境的所有有害影響降至低。
比較了兩個(gè)同基因系Kitaake（WT）及其基因改造品系CIPK2（RC），它們以較低的氮素利用效率（NUE）過(guò)量表達(dá)了鈣調(diào)磷酸酶B樣相互作用蛋白激酶2（OsCIPK2）的基因，并在低溫下對(duì)其生長(zhǎng)發(fā)育進(jìn)行了比較。與正常水平的氮相比。NUE是一個(gè)復(fù)雜的性狀，主要與植物從土壤中提取，吸收和循環(huán)利用N的效率有關(guān)。使用高通量Illumina Miseq 16S rRNA測(cè)序分析了微生物種群，發(fā)現(xiàn)帶有CIPK2的RC在水稻根部特異表達(dá)，不僅在不使用氮肥（LN）的情況下表現(xiàn)更好，而且增加了水稻根際區(qū)室細(xì)菌群落的多樣性（根際，根際和內(nèi)球）。
有益菌門(mén)的相對(duì)豐度增加，已知它可以促進(jìn)根際土壤中氮的循環(huán)和轉(zhuǎn)化。為了進(jìn)一步探索RC在LN下獲得更好性能的潛力，通過(guò)無(wú)創(chuàng)微測(cè)試技術(shù)（NMT）檢測(cè)了根尖中的離子通量。我們發(fā)現(xiàn)，與野生型相比，RC在LN下可以吸收更多的Ca2+和NO3-。后，與WT相比，RC植物在LN下表現(xiàn)出更好的根和莖生長(zhǎng)，并增加了產(chǎn)量和氮素吸收，而在正常氮（NN）處理下，兩個(gè)水稻品系的生長(zhǎng)沒(méi)有顯著差異。
通過(guò)研究與NUE相關(guān)的水稻分子，生理和化學(xué)參數(shù)，我們能夠獲得初步結(jié)果，處理過(guò)表達(dá)OsCIPK2的水稻品系。研究結(jié)果為進(jìn)一步研究水稻從土壤中吸收和利用氮以及與微生物群落的相互作用奠定了基礎(chǔ)。
結(jié)果表明：LN（low nitrogen）處理時(shí)，RC（過(guò)表達(dá)型）根系的Ca2+吸收顯著增加，而NN（normal nitrogen）處理時(shí)無(wú)顯著差異；LN處理時(shí)，RC根系的NO3-吸收增加，而NN條件下RC和WT（野生型）根系的NO3-表現(xiàn)為外排，且二者并無(wú)顯著差異；至于NH4+，RC和WT均表現(xiàn)為外排，且無(wú)顯著差異。這說(shuō)明，LN條件下，根系CIPK2基因過(guò)表達(dá)促進(jìn)了根系吸收NO3-，與RC根際的硝化作用增加是符合的。