Announcement: From today, the original "imOmics Essence Express" was revised to " Xuyue Living Research Communication ", referred to as " Xuyue Communication ", once a week. In addition to introducing the latest NMT literature, there are experimental design, equipment operation, data analysis, article writing, review and review questions... more dry goods, waiting for you to come.
NISC Document Number: C2015-036
Research using equipment
NMT Physiological Tester NMT Physiolyzer ®
Nitrate is the main nitrogen source for cereal crops, so mastering nitrate signaling in cereals is critical to improving nitrogen use efficiency. Although several regulators of nitrate signaling have been identified in Arabidopsis, they have not been found in cereals.
The Tong Yiping Group of the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences published an article on Plant Physiology entitled "The Nitrate-Inducible NAC Transcription Factor TaNAC2-5A ControlsNitrate Response and Increases Wheat Yield", which focuses on nitrate transport in cereals. reaction.
Overexpression of wild-type maize under low nitrogen conditions roots NO 3 - flow detection. A positive value indicates absorption.
The research team isolated a nitro-induced cereal-specific NAM, ATAF and CUC (NAC) transcription factor TaNAC2-5A from wheat. Chromatin immunoprecipitation assays showed that TaNAC2-5A binds directly to the promoter region of the nitrate transporter and glutamine synthetase encoding genes. Overexpression of TaNAC2-5A in wheat promotes root growth and nitrate uptake rate, thereby increasing the ability of roots to absorb nitrogen.
Utilization NMT vivo physiological detector Physiolyzer® based NMT technology (Non-invasive Micro-testTechnology, NMT) detects the root NO 3 - stream, we found NO 3 transfected plant genotypes roots - The rate of absorption was significantly higher than the wild type This provides the most direct physiological evidence for TaNAC2-5A to promote NO 3 - uptake in crop roots.
On June 4, 2018, Xuyue Research Institute went to the Institute of Genetics of the Chinese Academy of Sciences to conduct training on the application of non-invasive micro-test technology. The teachers and classmates at the scene raised several representative problems in the non-injury detection of potassium absorption of plants.
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First, why can't I detect potassium nutrient absorption?
Hunger treatment
· The primary concern of nutrient absorption experiments such as nitrogen (N) and potassium (K). N starvation, K starvation time, reference literature, generally 2-7 days (C2014-022, want the full text of the literature, please leave a message at the bottom of your mailbox and literature number).
· Some teachers may be reluctant to accept hunger treatment, and believe that there is another coercive factor "hunger stress" for their experiments. Of course, from a rigorous point of view, you can set up both hungry and non-starved groups at the same time with acceptable workload.
What I want to say is that before NMT appeared, the evidence for N and K absorption by plants mainly comes from the determination of N and K content, such as ICP-MS or isotope mass spectrometry. These detection methods are not real-time detection, but belong to “ The end point method, that is, detecting a "starting point" value, and then detecting an "end point" value, the difference between the two is the amount of plant absorption of N, K. But in fact, after the emergence of NMT, the test found that nutrients are not absorbed at all times (NMT is real-time monitoring), and for plant roots, not every part will absorb (NMT is micron-accurate) Fixed point detection). Affected by traditional technology, it is necessary to give the majority of researchers a certain amount of time to accept these new results. NMT found that the real-time absorption signals of plants on N and K are very weak, so it is necessary to amplify this process by means of "starvation" to observe the signal difference between groups.
2. Seedling age
· The change of ion current is an early event in plant physiology, that is, when the sample has no macroscopic difference, the ion current has begun to change. Conversely, when changes are already noticeable on a macroscopic scale, the difference in ion current may not be detected. Therefore, there is a time difference between the conventional experimental time point and the time point of the ion current. Generally, the ion current detection is higher than the conventional experimental time point.
· According to experience, Arabidopsis (C2011-003), rice (C2014-022), wheat, tomato and other herbs, the size is generally within 3 weeks. If it is too large, for example more than one month, the ion current signal may be weak.
· In particular, some teachers will tell me that other methods (ICP-MS) have found that when the seedling size is 30 days, you can see the difference in potassium content, or directly tell me that I hope to be consistent with other experimental conditions in the previous period ( Other experiments have used 30 days of seedlings). You can try it, but why don't I recommend you use too much seedlings for reasons like above.
3. Detection fluid concentration
According to experience, in many cases, the high K environment will inhibit the absorption of K by the roots. Therefore, the K concentration in the test solution is generally set to 0.1 mM to 0.5 mM. However, some experiments require detection at different K concentrations, which can be set by yourself.
4. Detection site
In “Hunger Treatmentâ€, we have already mentioned that not every part of the plant roots absorbs K. The reference finds that there is a meristematic zone at a distance of 100 μm from the root tip, which has obvious K absorption (C2011-003); It is a mature area (C2014-024) at a distance of 1.4 cm from the root tip. Therefore, at the beginning of the experiment, you need to scan all the roots of your sample ("sweeping root") to determine the most obvious site of K absorption signal, and then determine the spot.
NH 4 + and NO 3 - root results of poplar roots (C2014-025)
Second, the signal fluctuation is large when detecting K +
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1. Allow the sample to "wait" for a while before testing
Solution: The first few samples of the experiment were tested for a while and the time required for stabilization (balance phase) was explored.
The sample enters the test solution from the culture solution and needs to be adapted. So let the sample "stand" for a while in the test solution, which may be 5 minutes or 10 minutes, and then load the test. In this way, the effects of signal fluctuations caused by previous environmental changes can be ruled out. For specific reasons, watch the video "NMT Experimental Design and Mapping Training".
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2. Volatility signals may be important discoveries
Please refer to the blog post of Xu Yue, co-founder of modern non-damage micro-test technology. "The Nobel Prize Opportunity: How to understand and use NMT data? 》
This is a teacher and the Xunyue engineer who inadvertently recorded the data while checking out the gap and going out for lunch together. See what is the law (the yellow line is recorded before they eat)?
Third, the reviewer questioned whether the K + sensor detected K + in the end .
Xu Yue, co-founder of modern non-invasive micro-test technology, teaches you how to respond to the reviewer's questioning "How to determine the NMT measured is K + flow | Reviewer's question answer"
As of May 2018, a total of 216 NMT-related SCI articles were published by domestic scholars, with a total impact factor of 846.033 .
Note: The technical names of SIET, MIFE, SVET, SPET, etc. have been unified into Non-invasive Micro-test Technology, the Chinese name is “non-invasive micro-test technologyâ€, referred to as NMT.
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