CN109541133B - Early nondestructive identification method for nitrogen absorption capacity of tea tree - Google Patents
Early nondestructive identification method for nitrogen absorption capacity of tea tree Download PDFInfo
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Abstract
An early nondestructive identification method for nitrogen absorption capacity of tea trees, belonging to the technical field of tea tree cultivation. The method comprises the following steps: 1) selecting and cutting branches of the tea tree cuttings, and selecting and pretreating root systems; 2) fixing a non-invasive micro-measuring root system; 3) selecting a test solution; 4) correcting a non-destructive micrometering system; 5) surface NH of root system4 +And NO3 ‑Measuring the ion flow rate; 6) evaluating the nitrogen absorption capacity of the tea trees; 7) and (5) replanting the cutting seedlings. According to the early nondestructive identification method for the nitrogen absorption capacity of the tea trees, the tea tree resources are subjected to sand culture cuttage for a period of time, then the tea seedlings are taken out after root connection, and the root systems of the tea seedlings are subjected to nondestructive micro-measurement of the flow velocity of nitrate ions and ammonium ions so as to evaluate the nitrogen absorption capacity of the tea tree resources.
Description
Technical Field
The invention belongs to the technical field of tea tree cultivation, and particularly relates to an early nondestructive identification method for nitrogen absorption capacity of tea trees.
Background
The tea tree takes fresh leaves as a harvest object, the demand for nitrogen fertilizer is large, and a large number of researches show that the nitrogen fertilizer has an obvious yield increase effect on the tea. However, the problem of soil nutrient impoverishment of the tea garden is relieved by simple fertilization, so that the input cost of the tea industry and the potential environmental risk brought by the fertilizer are increased, and the sustainable development requirement of the tea industry is not met. Therefore, the method has important significance in breeding nitrogen-efficient tea plant varieties. The tea tree is a perennial economic crop, and the time for selecting improved varieties is long. In conclusion, the development of the rapid and effective early nondestructive identification technology for the nitrogen absorption capacity of the tea trees can not only greatly shorten the breeding time of the nitrogen-efficient tea tree variety, but also completely reserve the bred tea tree variety for later field verification. At present, the early nondestructive identification technology related to the nitrogen absorption capacity of tea trees is still blank.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to design a technical scheme of an early nondestructive identification method for the nitrogen absorption capacity of tea trees.
The early nondestructive identification method for the nitrogen absorption capacity of the tea trees is characterized by comprising the following steps:
1) selecting and cutting branches of the tea tree cutting slips, and selecting and pretreating root systems: respectively performing spike-strip maintenance and cuttage on the variety to be identified and the control variety, and cutting off the root system to be detected for later use after 2-4 cm of fresh tender roots grow out of the cuttage seedlings;
2) non-invasive microassay root fixation: fixing the root system to be tested at the bottom of the test tray;
3) selecting a test solution: adopts 0.08-0.12 mM CaCl20.25-0.35 mM morpholine ethanesulfonic acid MES and different concentrations of NH4NO3Mixing to obtain test solution, adjusting pH of the test solution to 6.3-6.8 with 0.5 mM Tris solution of Tris (hydroxymethyl) aminomethane (NH)4NO3Concentration test designs 0.1, 0.5, 1, 5, 8 mM concentration gradient, covering the concentration gradient of low nitrogen, medium nitrogen and high nitrogen of tea tree;
4) correcting a non-destructive micrometering system: before each test or probe replacement, correcting the non-destructive micro-test system by adopting a three-point method, and selecting NH4NO3The concentration is 0.1, 1, 8 mM;
5) surface NH of root system4 +And NO3 -Ion flow rate measurement: selecting a smooth surface locus of a root system in a mature area for testing,testing each locus for 2-4 min, selecting 3 loci for one root system, taking the average value as the flow rate value of one root after the flow rate is measured, and selecting 6 roots as biological repetition in each test;
6) evaluating the nitrogen absorption capacity of the tea trees: according to NH at different concentrations4 +And NO3 -Ion flow velocity, calculating total nitrogen flow velocity, solving absorption kinetic parameters of total nitrogen by adopting a Mie's kinetic equation, comparing the absorption kinetic parameters of the identified variety and the comparison variety, and evaluating the nitrogen absorption capacity of the identified variety;
7) and (5) replanting the cutting seedlings.
The early nondestructive identification method for nitrogen absorption capacity of tea trees is characterized by specifically comprising the following steps in the step 1):
a. selecting the cutting branches of the tea trees and carrying out sand culture cutting: respectively keeping spike strips of the variety to be identified and the control variety without picking spring tea of the tea tree, cutting off the tea tree branches with green stem sections after the tea tree branches become hard in the early 3 months, and then cutting the branches into 2.5-3.5 cm-long cuttings with one bud and one leaf; washing and fully airing sand used for sand culture, paving the sand in the field, wherein the thickness of the sand is 15-25cm, selecting rainy days, cutting the cut cutting slips into the sand, irrigating the sand after cutting, keeping 78-83% of the field water capacity, building a sunshade frame, and shading the sun by using two layers of sunshade nets;
b. selecting and pretreating a non-invasive micro-measuring root system: carefully taking out the root system from sand after the root system of the cutting seedling grows out, cleaning the sand on the surface of the root system with pure water, culturing in a tea tree nutrient solution until 2-4 cm of fresh tender root grows out, shearing the root system to be tested, and placing in a test solution for ion balance and stabilization for 5-15 min, preferably 10 min.
The early nondestructive identification method for nitrogen absorption capacity of tea trees is characterized by specifically comprising the following steps in the step 2): one end of the root system to be tested is clamped by two long and narrow filter papers and is placed in a test disc, and the two ends of each filter paper are respectively fixed by 1 flat-bottom small square stone of 1 multiplied by 1 cm, so that the root system to be tested is not clung to the bottom of the test disc, and the probe tip is prevented from being damaged in the test process.
The early nondestructive identification method for nitrogen absorption capacity of tea trees is characterized by comprising the following steps of 3): with 0.1 mM CaCl20.3 mM morpholine ethanesulfonic acid MES and different concentrations of NH4NO3As a test solution, the pH of the test solution was adjusted to 6.5 with a 0.5 mM Tris solution of Tris (hydroxymethyl) aminomethane.
The early nondestructive identification method for the nitrogen absorption capacity of the tea trees is characterized in that the tea tree nutrient solution is prepared by mixing the following substances: 1 mM NH4NO3、0.07 mM KH2PO4、0.3 mM K2SO4、0.67 mM MgSO4·7H2O、0.53 mM CaCl2·2H2O、 0.035 mM Al2(SO4)3·18H2O、7μM H3BO4 、1μM MnSO4·H2O、0.67μM ZnSO4·7H2O 、0.13μM CuSO4·5H2O、0.047μM (NH4)6Mo7O24·4H2O, 4.2. mu.M EDTA-Fe, balance purified water, adjusted to pH 5.0 with 0.01 mM NaOH.
The tea tree nutrient solution and the test solution are prepared by the tea research institute of Chinese academy of agricultural sciences.
According to the early nondestructive identification method for the nitrogen absorption capacity of the tea trees, the tea tree resources are subjected to sand culture cuttage for a period of time, then the tea seedlings are taken out after root connection, and the root systems of the tea seedlings are subjected to nondestructive micro-measurement of the flow velocity of nitrate ions and ammonium ions so as to evaluate the nitrogen absorption capacity of the tea tree resources.
Detailed Description
The present invention is further illustrated by the following specific examples.
Example 1:
1) selecting the cutting branches of the tea trees and carrying out sand culture cutting: selecting Longjing 43 as a high-nitrogen high-efficiency tea tree variety contrast, using Wuniao as a low-nitrogen high-efficiency tea tree contrast variety, using middle tea 302 as a variety to be identified, keeping spike strips of the variety to be identified (the middle tea 302) and the contrast variety (the Longjing 43 and the Wuniao) respectively for culture, cutting off tea tree branches with green stem sections after the tea tree branches become hard in the beginning of 3 months, and then cutting the branches into 2.5-3.5 cm long cuttings with one bud and one leaf. And washing and fully airing sand adopted by sand culture, and paving the sand in a field (the thickness is 20 cm). And (4) selecting rainy days, and cutting the cut cutting slips into sand. And after the cuttage is finished, irrigating the seedlings, and keeping 80% of the field water capacity. Building a sunshade frame, and shading with two layers of sunshade nets;
2) selecting and pretreating a non-invasive micro-measuring root system: carefully taking out the root system from sand after the root system of the cutting seedling grows out, cleaning the sand on the surface of the root system by using pure water, culturing in a tea tree nutrient solution until 2-4 cm of fresh tender roots grow out, shearing the root system to be detected, and placing in the solution to be detected for ion balance and stabilization for 10 min;
3) non-invasive microassay root fixation: clamping one end of a root system to be tested by two long and narrow filter papers, putting the clamped end into a test disc, and fixing two ends of each filter paper by 1 flat-bottomed diamond of 1 multiplied by 1 cm respectively, so that the root system to be tested is not attached to the bottom of the test disc, and a probe tip is prevented from being damaged in the test process;
4) selecting a test solution: with 0.1 mM CaCl20.3 mM MES and varying concentrations of NH4NO3As a test solution, the pH of the test solution was adjusted to 6.5, NH with 0.5 mM Tris solution4NO3Concentration test designs 0.1, 0.5, 1, 5, 8 mM concentration gradient, covering the concentration gradient of low nitrogen, medium nitrogen and high nitrogen of tea tree;
5) correcting a non-destructive micrometering system: before each test or probe replacement, correcting the non-destructive micro-test system by adopting a three-point method, and selecting NH4NO3The concentration is 0.1, 1, 8 mM;
6) surface NH of root system4 +And NO3 -Ion flow rate measurement: selecting smooth surface sites of the root system in a mature area for testing, wherein each site is tested for 3 min, 3 sites are selected for one root system, an average value is taken as a flow rate value of one root after flow rate measurement, and 6 roots are selected for biological repetition in each test;
7) evaluating the nitrogen absorption capacity of the tea trees: according to NH at different concentrations4 +And NO3 -Ion flow velocity, calculating total nitrogen flow velocity, solving absorption kinetic parameters of total nitrogen by adopting a Mie's kinetic equation, comparing the absorption kinetic parameters of the Chinese tea 302 and the reference varieties of the dragon well 43 and the Wuniu early, and finding through investigation that the Chinese tea 302 is larger than the dragon well 43 for Vmax value and the Chinese tea 302 is equivalent to the Wuniu early for alpha value; thus, the nitrogen absorbing capacity of middle tea 302 was identified as double highly potent.
8) Replanting the cutting seedling: and after the test is finished, the taken cutting seedlings are inserted back into the sand again for later test.
As can be seen from Table 1, the Vmax value of tea 302 in the tea variety to be identified is obviously superior to that of the high-nitrogen high-efficiency control variety Longjing 43, and the alpha value is equivalent to that of the low-nitrogen high-efficiency control variety Wuniu early. Thus, the nitrogen absorbing capacity of middle tea 302 was identified as double highly potent.
In step 3) of the above examples, 0.08 mM CaCl2, 0.25 mM MES morpholine ethanesulfonate and different NH concentrations were used as test solutions4NO3Mixing, and adjusting the pH of the test solution to 6.3 by using 0.5 mM Tris solution; or the test solution adopts 0.12 mM CaCl2, 0.35 mM morpholine ethanesulfonic acid MES and NH with different concentrations4NO3Mixing, and adjusting the pH of the test solution to 6.8 by using 0.5 mM Tris solution; the technical effects of the present invention can be obtained in the same manner as in example 1.
Claims (4)
1. An early nondestructive identification method for nitrogen absorption capacity of tea trees is characterized by comprising the following steps:
1) selecting and cutting branches of the tea tree cutting slips, and selecting and pretreating root systems: selecting Longjing 43 as a high-nitrogen high-efficiency tea tree variety contrast, taking Wuniu early as a low-nitrogen high-efficiency tea tree contrast variety, respectively performing scion stocking and cuttage on the variety to be identified and the contrast variety, and cutting off a root system to be detected after a cuttage seedling grows to 2-4 cm of fresh tender roots for later use;
the step 1) specifically comprises the following steps:
a. selecting the cutting branches of the tea trees and carrying out sand culture cutting: respectively keeping spike strips of the variety to be identified and the control variety without picking spring tea of the tea tree, cutting off the tea tree branches with green stem sections after the tea tree branches become hard in the early 3 months, and then cutting the branches into 2.5-3.5 cm-long cuttings with one bud and one leaf; washing and fully airing sand used for sand culture, paving the sand in the field, wherein the thickness of the sand is 15-25cm, selecting rainy days, cutting the cut cutting slips into the sand, irrigating the sand after cutting, keeping 78-83% of the field water capacity, building a sunshade frame, and shading the sun by using two layers of sunshade nets;
b. selecting and pretreating a non-invasive micro-measuring root system: carefully taking out the root system from sand after the root system of the cutting seedling grows out, cleaning the sand on the surface of the root system by using pure water, culturing in a tea tree nutrient solution until 2-4 cm of fresh tender roots grow out, shearing the root system to be tested, and placing in a test solution for ion balance and stabilization for 5-15 min;
2) non-invasive microassay root fixation: fixing the root system to be tested at the bottom of the test tray;
3) selecting a test solution: adopts 0.08-0.12 mM CaCl20.25-0.35 mM morpholine ethanesulfonic acid MES and different concentrations of NH4NO3Mixing to obtain test solution, adjusting pH of the test solution to 6.3-6.8 with 0.5 mM Tris solution of Tris (hydroxymethyl) aminomethane (NH)4NO3Concentration test designs 0.1, 0.5, 1, 5, 8 mM concentration gradient, covering the concentration gradient of low nitrogen, medium nitrogen and high nitrogen of tea tree;
4) correcting a non-destructive micrometering system: before each test or probe replacement, correcting the non-destructive micro-test system by adopting a three-point method, and selecting NH4NO3The concentration is 0.1, 1, 8 mM;
5) surface NH of root system4 +And NO3 -Ion flow rate measurement: selecting smooth surface sites in the mature region, testing for 2-4 min at each site, selecting 3 sites for one root system, measuring flow rate, averaging to obtain flow rate value of one root, and selecting 6 roots as flow rate value for each testBiological repetition;
6) evaluating the nitrogen absorption capacity of the tea trees: according to NH at different concentrations4 +And NO3 -Ion flow velocity, calculating total nitrogen flow velocity, solving absorption kinetic parameters of total nitrogen by adopting a Mie's kinetic equation, comparing the absorption kinetic parameters of the identified variety and the comparison variety, and evaluating the nitrogen absorption capacity of the identified variety;
7) and (5) replanting the cutting seedlings.
2. The method for the early non-destructive evaluation of nitrogen absorbing capacity of tea trees as claimed in claim 1, wherein the step 2) comprises the following steps: one end of the root system to be tested is clamped by two long and narrow filter papers and is placed in a test disc, and the two ends of each filter paper are respectively fixed by 1 flat-bottom small square stone of 1 multiplied by 1 cm, so that the root system to be tested is not clung to the bottom of the test disc, and the probe tip is prevented from being damaged in the test process.
3. The method for the early non-destructive evaluation of nitrogen uptake capacity of tea plant as claimed in claim 1, wherein in step 3): with 0.1 mM CaCl20.3 mM morpholine ethanesulfonic acid MES and different concentrations of NH4NO3As a test solution, the pH of the test solution was adjusted to 6.5 with a 0.5 mM Tris solution of Tris (hydroxymethyl) aminomethane.
4. The method of claim 1, wherein the tea plant nutrient solution is prepared by mixing: 1 mM NH4NO3、0.07 mM KH2PO4、0.3 mM K2SO4、0.67 mM MgSO4·7H2O、0.53 mM CaCl2·2H2O、 0.035 mM Al2(SO4)3·18H2O、7μM H3BO4 、1μM MnSO4·H2O、0.67μM ZnSO4·7H2O 、0.13μM CuSO4·5H2O、0.047μM (NH4)6Mo7O24·4H2O, 4.2. mu.M EDTA-Fe, the remainderThe amount was purified water, and the pH was adjusted to 5.0 with 0.01 mM NaOH.
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