CN109315286B - Method for screening corn genotype for improving nitrogen efficiency of corn/soybean intercropping system - Google Patents
Method for screening corn genotype for improving nitrogen efficiency of corn/soybean intercropping system Download PDFInfo
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Abstract
The invention discloses a method for screening and improving corn genotype of nitrogen efficiency of a corn/soybean intercropping system, belonging to the technical field of agricultural planting, and the technical scheme is as follows: the corn genotypes for improving the nitrogen efficiency of the corn/soybean intercropping system are screened out by screening the corn varieties with deep roots, strong capability of secreting flavonoids compounds from the roots and high system yield which are suitable for being combined with the soybean intercropping, then dividing the corn varieties into the corn varieties with deep roots and shallow roots and the soybean intercropping, and analyzing the physiological mechanism of the nitrogen efficiency difference of the corn/soybean intercropping system. The invention researches the underground interaction mechanism of the single cropping system and the influence on the nitrogen fixation efficiency of the intercropping soybeans by screening the corn varieties with different root system configurations and root exudates, clearly defines the root system morphological physiological characteristics of the corn varieties suitable for the corn/soybean intercropping system and can improve the overall nitrogen efficiency of the intercropping system.
Description
Technical Field
The invention belongs to the technical field of agricultural planting, and particularly relates to a technology for improving nitrogen efficiency of a corn/soybean intercropping system by screening corn varieties with different root system characteristics.
Background
The corn and soybean intercropping is a typical configuration representing the intercropping of high and short stalk crops and standing grain bean crops, and is a dry land planting mode with strong adaptability. The jade/bean mode is distributed all over the country before the country is built, is a novel efficient multi-cropping interplanting mode for dry land which is rapidly developed in the southwest area of China in recent years, and is an important measure for increasing both production and income, increasing the biological diversity of a farmland ecosystem, improving the utilization rate of nitrogen and reducing the using amount of a chemical fertilizer. The popularization area of the wheat-jade-bean planting mode in the Sichuan province in 2013 breaks through 40 ten thousand hectares. The advantages of intercropping yield are completely determined by the interspecific interaction of root systems of underground parts, the reasonable root configuration of gramineous crops and key root exudates play an important role in strengthening nodulation and nitrogen fixation of leguminous crops, and the method is a main way for efficiently utilizing nitrogen of jade/bean systems.
Crop combination and proper variety selection in intercropping are the key to the success of intercropping. The nitrogen acquisition capacity and the root secretion capacity of different corn varieties have obvious difference, which may affect the nitrogen fixation effect of soybeans in an intercropping system and finally affect the efficient utilization of nitrogen in the whole production system. However, there is currently no basis for selection of maize varieties in intercropping systems.
Disclosure of Invention
In order to overcome the defects, the inventor of the invention continuously reforms and innovates through long-term exploration and trial and multiple experiments and efforts, and provides a method for improving the nitrogen efficiency of a corn/soybean intercropping system by screening root system characteristic difference.
The specific technical scheme is as follows: a method for screening corn genotypes for improving nitrogen efficiency of a corn/soybean intercropping system is characterized by comprising the following steps: the method comprises the following operations:
A. screening a genotype corn variety which is suitable for being combined with soybean intercropping, has strong capability of secreting flavonoid compounds from roots and high system yield, and comprises the following specific operations:
a 1: selecting a corn variety to be screened,
a 2: setting planting areas with the same nitrogen fertilizer level, then dividing the planting areas into single cropping mode and intercropping mode for planting, planting corn and soybean in the corresponding areas,
a3 fertilization: respectively applying nitrogen fertilizer to the corn strips before sowing and in the jointing stage, applying start fertilizer to the soybean strips before sowing, inoculating rhizobia before sowing,
a4, digging roots of corns and sampling in the spinning period, photographing the root systems of the samples, analyzing the root systems by using DIRT software, simultaneously taking partial samples of the ground part of the root systems, digesting and measuring nitrogen,
a5, harvesting and measuring yield in the mature period, sampling soybeans in the mature period, picking root nodules, then counting, air-drying, weighing, calculating the biological nitrogen fixation amount by using a 15N natural abundance method,
a6, screening out the corn genotype which is most suitable for the combination with soybean intercropping, has strong capability of secreting flavonoids from roots and high system yield through an indoor water culture test;
B. utilizes deep-root and shallow-root maize varieties and soybean intercropping to analyze the physiological mechanism of nitrogen efficiency difference of the maize/soybean intercropping system so as to screen out maize genotypes for improving the nitrogen efficiency of the maize/soybean intercropping system,
b1, intercropping the corn varieties with different root systems screened from the a6 with soybeans respectively, setting two intercropping modes,
b2 setting three different nitrogen fertilizer horizontal planting areas with different heights, setting two intercropping modes in each nitrogen fertilizer horizontal planting area,
b 3: in the middle-high nitrogen fertilizer horizontal area, the corn stripe nitrogen fertilizer is applied twice before sowing and in the jointing stage, the soybean stripe is applied with a start fertilizer before sowing, and rhizobium is inoculated before soybean sowing;
b4 digging root of corn in the stage of jointing, large flare, silk-spitting and mature, sampling, taking the sample on the ground, digesting and measuring nitrogen, detecting the content of nitrate nitrogen and ammonium nitrogen in soil,
b5, finally harvesting and measuring yield; meanwhile, an LI-6400 type photosynthesis system is adopted in the morning of a sunny day at 9: 00-11: selecting leaves of corn between 00 to determine net photosynthetic rate;
b6, sampling soybeans in a mature period, picking visible root nodules, counting, air-drying, weighing, and calculating the biological nitrogen fixation amount by using a 15N natural abundance method; thus determining the corn genotype capable of improving the nitrogen efficiency of the corn/soybean intercropping system according to the nitrogen fixation amount.
According to the method for screening the corn genotype capable of improving the nitrogen efficiency of the corn/soybean intercropping system, the further preferable technical scheme is that the nitrogen fertilizer level in a2 is 150kg/hm2。
According to the method for screening and improving the nitrogen efficiency of the corn/soybean intercropping system, the preferable technical scheme is that in the a2, the corn pit distance of the single cropping standard is 25cm, the line distance is 40cm multiplied by 120cm, the wide and narrow lines are arranged, the line distance between the corn and the soybean is 30cm, the plant distance between the soybean is 25cm, and 6-8 grains are dibbled in holes.
The method for screening the corn genotype capable of improving the nitrogen efficiency of the corn/soybean intercropping system has the further preferable technical scheme that: the planting density in the planting area is 5 ten thousand plants/hm2。
According to the method for screening and improving the nitrogen efficiency of the corn/soybean intercropping system, the further preferable technical scheme is that nitrogen fertilizers are respectively applied before the corn is sowed in a3 in a strip and in the jointing stage, and the application amount of the nitrogen fertilizers is the same.
According to the method for screening and improving the corn genotype of the nitrogen efficiency of the corn/soybean intercropping system, the preferable technical scheme is that a6 water culture experiment specifically comprises the steps of carrying out water culture on a corn variety, passing culture solution in a pot through a macroporous resin column after 3 weeks of culture to collect root exudates, then eluting flavonoids compounds in the resin with 85% alcohol, concentrating the collected eluent through a nitrogen blowing instrument, and measuring the flavonoids compounds by using HPLC (high performance liquid chromatography), wherein the measurement wavelength is 280 nm.
According to the method for screening the corn genotype capable of improving the nitrogen efficiency of the corn/soybean intercropping system, a further preferable technical scheme is that when the content of nitrate nitrogen and ammonium nitrogen in soil is detected in b4, soil samples are taken every 30cm, and the soil sampling depth is 0-90 cm.
According to the method for screening the maize genotype for improving the nitrogen efficiency of the maize/soybean intercropping system, a further preferable technical scheme is that during the soybean maturation period sampling in b6, iron is used for lifting and digging soil to take out 5 broad beans and place the whole broad beans on kraft paper to ensure that all root nodules are taken out, soil on the roots is shaken off gently, the root nodules and the roots with the root nodules falling on the paper are collected, the shaken off soil is placed back to the original sampling position, and the sample is placed in a 100-mesh sieve for running water and washing after being taken back.
According to the method for screening and improving the nitrogen efficiency of the corn/soybean intercropping system, a further preferable technical scheme is that two intercropping modes in b1 are adopted, wherein the row spacing of the corn and the soybean in the first intercropping mode is 30cm, the row spacing of the corn and the soybean in the second intercropping mode is 40cm, the pit spacing of the corn cultivation specification is 25cm, the row spacing is 40cm multiplied by 120cm wide and narrow rows, the soybean plant spacing is 25cm, 6-8 grains are dibbled, and 5 plants are reserved in each hole during seedling setting.
According to the method for screening the corn genotype capable of improving the nitrogen efficiency of the corn/soybean intercropping system, the further preferable technical scheme is that the planting density in two intercropping modes in b1 is 5 ten thousand plants/hm2
The invention researches the underground interaction mechanism of the single cropping system and the influence on the nitrogen fixation efficiency of the intercropping soybeans by screening the corn varieties with different root system configurations and root exudates, clearly defines the root system morphological physiological characteristics of the corn varieties suitable for the corn/soybean intercropping system and can improve the overall nitrogen efficiency of the intercropping system.
Drawings
FIG. 1: in the examples, a schematic diagram of the planting pattern of corn/soybean intercropping is shown.
FIG. 2: the second schematic diagram of the planting mode of corn/soybean intercropping in the examples.
Detailed Description
The technical scheme of the invention is further described in detail through experiments by combining the specific practical environment.
Examples
This embodiment is accomplished primarily in two major steps.
Firstly, the method comprises the following steps: screening the corn genotype which is most suitable for combining with soybean intercropping, has strong capability of secreting flavonoid compounds from roots and high system yield, and is mainly completed by the following steps;
and (3) experimental design:
15-20 major maize varieties in Sichuan are selected in recent years, and 1 nitrogen fertilizer level (150 kg/hm) is set in the test2) The method comprises two planting modes of single cropping and intercropping (as shown in figure 1). Four replicates per treatment, cell area 16m2(3.2m is multiplied by 5m), the corn cultivation specification nest spacing is 25cm, the line spacing is wide and narrow lines (40cm is multiplied by 120cm), and the line spacing between corn and soybean is 30 cm. 2 rows are planted in each planting zone, and the planting density is 5 ten thousand plants/hm2. The distance between the soybean plants is 25cm, 6-8 soybean plants are dibbled in each hole, and 5 soybean plants are reserved in each hole during final singling. The corn stripe nitrogen fertilizer is applied twice (before sowing: the jointing period is 1: 1), and the soybean stripe nitrogen fertilizer is applied by 35kg/hm before sowing2And inoculating rhizobium before sowing soybean.
Measurement items and technical methods:
the corn is sampled by digging roots during the spinning period, and representative 3 plants are selected in each cell. Firstly, digging a soil column with the diameter of 35 cm and the depth of 30cm by taking the base part of a corn stem as a center, slightly shaking the root system of the corn after digging out, removing partial loosened soil by hands, and soaking the root in a mild detergent; finally, a small amount of soil still adhered to the root system is washed by a low-pressure water gun. After the root system is dried, taking a root system photograph, wherein the root system photograph method is referred to (Bucksch et al, 2014). An 80 cm square studio (DEEP brand, beijing white light camera, inc.) was used, a camera (SONYILCE 5100) was placed in the middle of the studio ceiling, the roots were dried and placed in the middle of the studio, and the roots were photographed and analyzed with DIRT software. Meanwhile, a sample of the overground part is taken, and the nitrogen is measured by digestion. Harvesting in the mature period and measuring yield. Sampling soybeans in a mature period, digging by a shovel about 0.60m deep to take out 5 broad beans of the whole plant, placing the broad beans on kraft paper to ensure that all root nodules are taken out, slightly shaking off soil on the roots, collecting the root nodules falling on the paper and the roots with the root nodules, and putting the shaken off soil back to the original sampling position. And after the sample is taken back, the sample is placed in a 100-mesh sieve for running water washing, and root nodules are picked up, counted, air-dried and weighed. And calculating the biological nitrogen fixation amount by using a 15N natural abundance method.
The indoor water culture test screens the genotype with strong capability of root secreting flavonoid compounds: culturing 15-20 corn varieties in water, culturing for 3 weeks, passing the culture solution in a pot through a macroporous resin column to collect root exudates, eluting flavonoids compounds in the resin with 85% alcohol, concentrating the collected eluate with a nitrogen blower, and measuring the flavonoids compounds with HPLC (high performance liquid chromatography) with a measurement wavelength of 280 nm.
And (3) combining the test results, screening out the corn genotype which is most suitable for the combination with soybean intercropping, has strong capability of secreting flavonoids from roots and high system yield.
II, secondly: the physiological mechanism of nitrogen efficiency difference of a corn/soybean intercropping system is analyzed by using deep-root and shallow-root corn genotypes and soybean intercropping.
And (3) experimental design:
and (3) intercropping the 4 corn genotypes (deep root 1, deep root 2, shallow root 1 and shallow root 2) with different root system sizes screened in the first step with soybeans. Test set 3 nitrogen fertilizer levels (0, 150, 300 kg/hm)2) Two intercropping modes are horizontally arranged for each nitrogen fertilizer, wherein the row spacing of the corn and the soybeans in the first intercropping mode is 30cm (shown in figure 1), and the row spacing of the corn and the soybeans in the second intercropping mode is 40cm (shown in figure 2). Four replicates per treatment, cell area 16m2(3.2m is multiplied by 5m), the corn cultivation specification nest spacing is 25cm, the row spacing is wide and narrow rows (40cm is multiplied by 120cm), 2 rows are planted in each planting zone, and the planting density is 5 ten thousand plants/hm2. The distance between the soybean plants is 25cm, 6-8 soybean plants are dibbled in each hole, and 5 soybean plants are reserved in each hole during final singling. At 150 and 300kg/hm2In the nitrogen fertilizer treatment, the corn stripe nitrogen fertilizer is applied twice (before sowing: the jointing period is 1: 1), and the soybean stripe nitrogen fertilizer is applied by 35kg/hm of starting fertilizer before sowing2And inoculating rhizobium before sowing soybean.
Measurement items and technical methods:
the root of the corn is dug and sampled in the jointing stage, the large-horn mouth stage, the silk-spitting stage and the mature stage, and the sampling method is the same as the content. Meanwhile, taking an overground part sample, digesting and measuring nitrogen, taking a soil sample every 30cm, taking a depth of 0-90cm, and detecting the content of nitrate nitrogen and ammonium nitrogen in the soil. And finally harvesting and measuring yield. Meanwhile, an LI-6400 type photosynthesis system (LI-COR, USA) is adopted in the morning of fine weather at 9: 00-11: leaf blade selection between 00 and 5 maize plants for determination of net photosynthetic Rate (. mu.mol CO 2/m)2/s)。
Soybeans were sampled at maturity. Lifting and digging by iron about 0.50m deep, taking out 5 broad beans of the whole plant, placing the broad beans on kraft paper to ensure that all the root nodules are taken out, lightly shaking off soil on the roots, collecting the root nodules and the roots with the root nodules which fall on the paper, and placing the shaken off soil back to the original sampling position. And after the sample is taken back, the sample is placed in a 100-mesh sieve for running water washing, and root nodules are picked up, counted, air-dried and weighed. And calculating the biological nitrogen fixation amount by using a 15N natural abundance method.
By combining the test results, the key corn root configuration and the key period of root system interaction for improving the nitrogen efficiency of the corn/soybean intercropping system are determined.
From the above-mentioned experimental result, combine the analysis data of the experiment and thus can screen out the corn genotype that improves nitrogen efficiency in the system that the maize soybean intercrops rationally, thus improve the efficiency of farming, improve the income of grower, increase economic benefits.
The test area selected by the embodiment is in Sichuan, and a person skilled in the art can modify the area into any area according to actual conditions, and perform corresponding tests by combining local actual environments.
Based on the above embodiments of the present invention, all other embodiments obtained by a person of ordinary skill in the art without any inventive work are within the scope of the present invention. Thus, the detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention.
Claims (10)
1. A method for screening corn genotypes for improving nitrogen efficiency of a corn/soybean intercropping system is characterized by comprising the following steps: the method comprises the following operations:
A. screening a genotype corn variety which is suitable for being combined with soybean intercropping, has strong capability of secreting flavonoid compounds from roots and high system yield, and comprises the following specific operations:
a 1: selecting a corn variety to be screened,
a 2: setting planting areas with the same nitrogen fertilizer level, then dividing the planting areas into single cropping mode and intercropping mode for planting, planting corn and soybean in the corresponding areas,
a3: fertilizing: respectively applying nitrogen fertilizer to the corn strips before sowing and in the jointing stage, applying start fertilizer to the soybean strips before sowing, inoculating rhizobia before sowing,
a4: digging roots of corn in the spinning period, taking a picture of the root system of the sample, analyzing the root system by using DIRT software, simultaneously taking a part of the sample on the ground of the root system, digesting and measuring nitrogen,
a5: harvesting and measuring yield in the mature period, sampling soybeans in the mature period, picking out root nodules, then counting, air-drying, weighing, calculating the biological nitrogen fixation amount by using a 15N natural abundance method,
a6: screening out a corn genotype which is most suitable for being combined with soybean intercropping, has strong capability of secreting flavonoid compounds from roots and high system yield through an indoor water culture test;
utilizes deep-root and shallow-root maize varieties and soybean intercropping to analyze the physiological mechanism of nitrogen efficiency difference of the maize/soybean intercropping system so as to screen out maize genotypes for improving the nitrogen efficiency of the maize/soybean intercropping system,
b1: the corn varieties with different root systems screened from a6 are intercropped with soybeans respectively, two intercropping modes are set,
b2: three different nitrogen fertilizer horizontal planting areas with different heights are arranged, each nitrogen fertilizer horizontal planting area is provided with two intercropping modes,
b 3: in the middle-high nitrogen fertilizer horizontal area, the corn stripe nitrogen fertilizer is applied twice before sowing and in the jointing stage, the soybean stripe is applied with a start fertilizer before sowing, and rhizobium is inoculated before soybean sowing;
b4: respectively digging roots of corns for sampling in the jointing stage, the large-horn mouth stage, the silk-throwing stage and the mature stage, simultaneously taking a sample on the ground, digesting and measuring nitrogen, detecting the content of nitrate nitrogen and ammonium nitrogen in soil,
b5: finally harvesting and measuring yield; meanwhile, an LI-6400 type photosynthesis system is adopted in the morning of a sunny day at 9: 00-11: selecting leaves of corn between 00 to determine net photosynthetic rate;
b6: sampling soybeans in a mature period, picking visible root nodules, counting, air-drying, weighing, and calculating the biological nitrogen fixation amount by using a 15N natural abundance method; thus determining the corn genotype capable of improving the nitrogen efficiency of the corn/soybean intercropping system according to the nitrogen fixation amount.
2. The method for screening corn genotypes for increased nitrogen efficiency in corn/soybean intercropping system according to claim 1, wherein the nitrogen fertilizer level in a2 is 150kg/hm2。
3. The method for screening the corn genotype capable of improving the nitrogen efficiency of the corn/soybean intercropping system according to claim 1, wherein in the a2, the row spacing of the corn with single cropping size is 25cm, the row spacing is 40cm x 120cm wide and narrow rows, the row spacing between the corn and the soybean is 30cm, the plant spacing between the soybean is 25cm, and 6-8 grains are dibbled in holes.
4. The method for screening corn genotypes for improving nitrogen efficiency of corn/soybean intercropping system according to claim 1, wherein the planting density in the planting area is 5 ten thousand plants/hm2。
5. The method for screening corn genotypes for improving nitrogen efficiency of corn/soybean intercropping system according to claim 1, wherein nitrogen fertilizer is applied separately in a3 before corn streak sowing and during jointing stage, and the nitrogen fertilizer is applied in the same amount.
6. The method for screening the corn genotype capable of improving the nitrogen efficiency of the corn/soybean intercropping system according to claim 1, wherein the water culture experiment in a6 is specifically to perform water culture on a corn variety, collect root exudates by passing culture solution in a pot through a macroporous resin column after 3 weeks of culture, elute flavonoids compounds in the resin with 85% alcohol, concentrate the collected eluate through a nitrogen blowing instrument, measure the flavonoids compounds by HPLC, and measure the wavelength at 280 nm.
7. The method for screening corn genotypes for improving nitrogen efficiency of corn/soybean intercropping system according to claim 1, wherein the soil nitrate nitrogen and ammonium nitrogen content is detected in b4, the soil depth is 0-90cm for every 30cm soil sample.
8. The method for screening maize genotypes for improved nitrogen efficiency in the maize/soybean intercropping system according to claim 1, wherein during soybean maturation period sampling in b6, the entire plant of broad bean 5 was removed by iron-lifting and earth-digging and placed on kraft paper to ensure all nodules were removed, the soil on the roots was gently shaken off, the nodules and roots with nodules that fell on the paper were collected and returned to the original sampling site, and the samples were rinsed in a 100 mesh sieve with running water after being retrieved.
9. The method for screening the maize genotypes for improving the nitrogen efficiency of the maize/soybean intercropping system according to claim 1, wherein two intercropping modes in b1 are provided, wherein the row spacing of the maize and the soybean in the first intercropping mode is 30cm, the row spacing of the maize and the soybean in the second intercropping mode is 40cm, the pit spacing of the maize cultivation specification is 25cm, the row spacing is 40cm x 120cm wide and narrow rows, the plant spacing of the soybean is 25cm, 6-8 grains are dibbled in the holes, and 5 plants are left in each hole during the final singling.
10. The method for screening corn genotype for improving nitrogen efficiency of corn/soybean intercropping system according to claim 1, wherein planting density in two intercropping modes of b1 is 5 ten thousand plants/hm2。
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