CN109593779B - Screening method and screening device for drought-enduring plants - Google Patents
Screening method and screening device for drought-enduring plants Download PDFInfo
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Abstract
The invention relates to the technical field of plant breeding, in particular to a screening method and a screening device for drought-enduring plants. Selecting a plurality of 2-year-old robust cutting seedlings or seedlings of commonly-used afforestation trees and shrubs, and measuring the initial root conditions and the seedling heights of the seedlings; simulating a desert drought environment in a simulation box, wherein the simulation box comprises a plurality of simulation areas; setting different rainfall degrees in different simulation areas, respectively planting plants in sand layers arranged at the bottoms of the simulation areas, simulating sunshine for 12-16h/d by using a fluorescent lamp at the illumination intensity of 4000-; selecting plant genes with good root system condition and good tree growth condition, screening and carrying out large-scale propagation. The method can effectively avoid the difference between the experimental environment and the field environment so as to screen the plants more suitable for the desert drought environment in the natural state.
Description
Technical Field
The invention relates to the technical field of plant breeding, in particular to a screening method and a screening device for drought-enduring plants.
Background
Currently, land desertification has become a global environmental problem and is also a great challenge for global plant growth. According to the 5 th report of China desertification and desertification conditions issued by the national forestry Bureau, the total area of the nationwide desertification land is 261.16 ten thousand km as long as 20142Accounts for 27.20 percent of the total area of the national soil, wherein the area of the Xinjiang desert is 107.06 km ten thousand2And accounts for 40.99% of the desert area in China. Xinjiang is the largest province of China desert area, and due to the complex landform, rare rainfall, obvious continental drought climate, and the influence of climate change and human factors, soil degradation, continuous expansion of desert area and situation are causedIs severe. The serious desertification of the land caused by the over utilization of the land or the lack of water resources is the most important harm for the social and economic development of arid areas. In order to reduce water consumption and transpiration area, desert plants are usually conifer species even without leaves, and only tender branches are used for photosynthesis, the species of the plants are few, and the shape is single, so that a drought-tolerant plant screening technology which can realize diversification of arbor species in desert areas and is configured for plants in desert arid and barren areas is needed.
Disclosure of Invention
The invention aims to solve the problems and provides a screening method and a screening device capable of screening out plants suitable for desert drought environments in natural states.
The technical scheme for solving the problems is to provide a screening method of drought-enduring plants, which comprises the following steps:
(1) plant sample preparation: selecting a plurality of 2-year-old robust cutting seedlings or seedlings of commonly-used afforestation trees and shrubs, and measuring the initial root conditions and the seedling heights of the seedlings;
(2) desert environment simulation: simulating a desert drought environment in a simulation box, wherein the simulation box comprises a plurality of simulation areas;
(3) drought tolerance screening: setting different rainfall degrees in different simulation areas, respectively planting plants in sand layers arranged at the bottoms of the simulation areas, simulating sunshine for 12-16h/d by using a fluorescent lamp at the illumination intensity of 4000-;
(4) gene screening: selecting plants with good root system conditions and good tree growth conditions, respectively extracting RNA of the plants, synthesizing full-length cDNA by using an RNA 5' end template conversion technology, and recovering large fragments of more than 500 bp; connecting the recovered product with pYES2.1 vector, transforming cDNA into Saccharomyces cerevisiae competent cells by electrotransformation, and culturing with uracil auxotroph culture medium to obtain yeast transformant; screening the yeast transformant in the gradient culture medium with the reduced water content, and selecting the yeast transformant with high drought tolerance to inoculate the yeast transformant in the YPGal culture medium for amplification culture; construction of plant expression vector by yeast transformant and its transfer into various plants
Preferably, the plant sample comprises evergreen arbor: dragon juniper, Japanese pine, Cinnamomum japonicum Thunb, Machilus thunbergii, Zingiber zerumbet Maxim, Ligustrum lucidum ait, evergreen Chinese wax and ilex chinensis Benth; also includes deciduous trees: alder, beech, Sophora japonica, Toona sinensis, Sapindus mukorossi and jujube tree; also includes evergreen shrubs: ramulus et folium Cacaliae Caudatae, Photinia fraseris, flos Rosae chinensis, ramulus Buxi Sinicae, Aleurites fordii, folium seu cortex Nerii, Ligustrum japonicum, and flos Hibisci Mutabilis; also includes deciduous shrubs: malus halliana, Morus alba and Lagerstroemia indica.
The invention also aims to provide a screening device for drought-enduring plants, which comprises a simulation box, wherein the top of the simulation box is provided with the fluorescent lamp, and the bottom of the simulation box is provided with the sandy soil layer; the simulation box is established in proper order into a plurality of simulation districts through a plurality of baffles in order to follow the horizontal direction with the simulation box inner chamber, the simulation box is equipped with the transparent plate of slope so that form the precipitation chamber between transparent plate and the fluorescent lamp, and makes the precipitation chamber in the different simulation districts vary in size, the transparent plate includes the closing plate and is equipped with the precipitation board of through-hole.
Preferably, be equipped with the vortex subassembly in the simulation case, the vortex subassembly is including transversely setting up the pivot in the simulation case, the pivot is equipped with horizontal vortex leaf, horizontal vortex leaf pass through the eccentric orfice with the pivot is connected.
Preferably, the spoiler assembly further comprises a vertical spoiler blade, the vertical spoiler blade comprises a shaft body and a connecting sleeve sleeved on the shaft body through an eccentric hole, and the connecting sleeve is provided with a blade; the shaft body with the pivot passes through umbelliform gear structural connection, umbelliform gear structure is including setting up the first umbelliform gear of pivot and setting up in the second umbelliform gear of shaft body, first umbelliform gear and the meshing of second umbelliform gear.
Preferably, the shaft body of the vertical spoiler blade is provided with a rolling bearing, and the rolling bearing is connected with a support piece arranged on the side wall of the partition plate or the simulation box.
Preferably, one end of the transparent plate is hinged to the inner wall of the simulation box, and the other end of the transparent plate is arranged on the arc-shaped wall of the simulation box in a sliding mode through a rubber sealing piece; the baffle includes expansion plate and lower expansion plate, the transparent plate sets up between upper expansion plate and the lower expansion plate and is connected with upper expansion plate and lower expansion plate respectively.
Preferably, the system also comprises a micro root chamber system, wherein the micro root chamber system comprises a micro root pipe arranged in a sandy soil layer, a micro CCD camera arranged outside the simulation box, a regulating and controlling handle, an image acquisition control device and a computer image processing and analyzing system.
The invention has the beneficial effects that:
set up simulation case simulation desert arid environment, carry out drought-enduring screening in the simulation incasement and can screen out the plant that is more suitable for arid environment under the natural state, effectively avoid experimental environment and the difference of on-the-spot environment, plant field planting for the later stage is screened provides good basis.
Drawings
FIG. 1 is a schematic diagram of a screening device for drought tolerant plants;
in the figure: the simulation box 100, the simulation area 101, the sand layer 102, the fluorescent lamp 103, the transparent plate 110, the rotating shaft 210, the horizontal spoiler 211, the vertical spoiler 220, the shaft body 221 and the micro root chamber system 300.
Detailed Description
The following are specific embodiments of the present invention and are further described with reference to the accompanying drawings, but the present invention is not limited to these embodiments.
A screening method of drought-enduring plants comprises the following steps:
(1) plant sample preparation: selecting a plurality of 2-year-old robust cutting seedlings or seedlings of commonly-used afforestation trees and shrubs, and measuring the initial root conditions and the seedling heights of the seedlings. In order to improve the diversity of plants planted on the spot in the arid desert area in the later period, more kinds of plants are selected as much as possible. Selecting evergreen trees such as juniper, Japanese cypress, sassafras prunifolia, photinia serrulata, Zingiber zerumbet, Ligustrum lucidum ait, evergreen Chinese wax, ilex chinensis and the like; deciduous trees such as alder, beech, Sophora japonica, Toona sinensis, Sapindus mukorossi, and jujube tree; evergreen shrubs such as Rhaphiolepis umbellata, Photinia fraseri, Rosa chinensis, Populus tremula, Aleurites fordii, Nerium oleander, Ligustrum japonicum, and flos Hibisci Mutabilis; deciduous shrubs such as Malus halliana, Morus alba, and Lagerstroemia indica.
(2) Desert environment simulation: the simulation of the desert drought environment is performed in a simulation box 100, and the simulation box 100 comprises a plurality of simulation areas 101. As shown in fig. 1, a fluorescent lamp 103 is arranged at the top of the simulation box 100, and a sand layer 102 is arranged at the bottom; simulation case 100 sets gradually a plurality of simulation districts 101 through a plurality of baffles in order to follow the horizontal direction with simulation case 100 inner chamber, and simulation case 100 is equipped with the transparent plate 110 of slope so that form the precipitation chamber between transparent plate 110 and the fluorescent lamp 103, and makes the precipitation chamber in the different simulation districts 101 vary in size, and transparent plate 110 includes the closing plate and is equipped with through-hole precipitation plate.
Because the precipitation chamber variation in size, then to the water delivery of precipitation intracavity and fill up the precipitation chamber, then take out from the closing plate and can make the precipitation degree inconsistent in each simulation area 101, obtain the experiment precipitation degree gradient simply to screen the drought tolerance of plant.
Meanwhile, in order to further adjust the precipitation degree, one end of the transparent plate 110 is hinged to the inner wall of the simulation box 100, and the other end is slidably arranged on the arc-shaped wall of the simulation box 100 through a rubber sealing element; the baffle includes expansion plate and lower expansion plate, and transparent plate 110 sets up between last expansion plate and the lower expansion plate and is connected with last expansion plate and lower expansion plate respectively, can change the capacity in whole precipitation chamber so through rotating transparent plate 110.
Desert environment still has the characteristics of many strong winds, many dust, still need simulate the wind resistance of plant in desert environment, because among the natural environment, the flow direction of wind presents irregular turbulent flow state, consequently is equipped with the vortex subassembly in the simulation case 100, and the vortex subassembly is including transversely setting up in the pivot 210 of simulation case 100, and pivot 210 is equipped with horizontal vortex leaf 211, and horizontal vortex leaf 211 passes through the eccentric orfice and is connected with pivot 210. The spoiler assembly further comprises a vertical spoiler blade 220, the vertical spoiler blade 220 comprises a shaft body 221 and a connecting sleeve sleeved on the shaft body 221 through an eccentric hole, and the connecting sleeve is provided with a blade. So as to simulate wind force in natural desert environment in a mixed mode of eccentric rotation around a horizontal shaft and eccentric rotation around a vertical shaft.
In addition, in order to achieve the energy saving effect, the shaft body 221 is connected with the rotating shaft 210 through a bevel gear structure, the bevel gear structure comprises a first bevel gear arranged on the rotating shaft 210 and a second bevel gear arranged on the shaft body 221, and the first bevel gear is meshed with the second bevel gear, so that the purpose of linkage is achieved through the same motor.
Meanwhile, in order to maintain the stability of the vertical spoiler 211, the shaft body 221 of the vertical spoiler 220 is provided with a rolling bearing connected with a support member provided at a sidewall of the barrier or the simulation box 100.
(3) Drought tolerance screening: setting different precipitation degrees in different simulation areas 101, respectively planting plants in sand layers 102 arranged at the bottoms of the simulation areas 101, simulating sunshine for 12-16h/d by using a fluorescent lamp 103 at the illumination intensity of 4000-.
The root system condition can be measured by adopting the micro root chamber system 300, the micro root chamber system 300 comprises micro root tubes arranged in a sand layer, a sand-soil mixing layer and a soil layer, a micro CCD camera, a regulating and controlling handle, an image acquisition control device and a computer image processing and analyzing system which are arranged outside a simulation box, and the detection mode not only avoids destructive, time-consuming and labor-consuming sampling processes, but also can directly carry out dynamic automatic measurement and analysis on the plant root system growing in the sand in situ and in a lossless manner.
(4) Gene screening: selecting plants with good root system conditions and good tree growth conditions, respectively extracting RNA of the plants, synthesizing full-length cDNA by using an RNA 5' end template conversion technology, and recovering large fragments of more than 500 bp; connecting the recovered product with pYES2.1 vector, transforming cDNA into Saccharomyces cerevisiae competent cells by electrotransformation, and culturing with uracil auxotroph culture medium to obtain yeast transformant; screening the yeast transformant in the gradient culture medium with the reduced water content, and selecting the yeast transformant with high drought tolerance to inoculate the yeast transformant in the YPGal culture medium for amplification culture; plant expression vectors were constructed from yeast transformants and transferred to various plants.
The technical method for converting the RNA 5' end template comprises the following steps: first strand cDNA synthesis was performed using a modified oligo G (dT) primer, oligonucleotide as a template for extension of mRNA5 ' end, and when reverse transcription reached mRNA5 ' end, the reverse transcription template-independent addition of several C's at first strand cDNA 3 ' end paired with several G's at oligonucleotide 3 ' end, thereby linking oligonucleotide to mRNA5 ' end. The reverse transcriptase jumps and reverse transcribes to the end of the oligonucleotide using the oligonucleotide as a template, and this jump often occurs in the eukaryotic mRNA cap structure-m 7GpppX, so that the reverse transcribed cDNA contains the entire 5' sequence of the mRNA and the complement of the oligonucleotide, followed by PCR to synthesize the second strand of cDNA. The cDNA clone contains the complete 5' non-coding region sequence of the corresponding mRNA molecule to the maximum extent, so that the full-length cDNA library of the salt-tolerant plant is constructed.
The simplest eukaryotic model organism, namely yeast, is used as a host of a plant exogenous gene, and because a yeast expression system is closer to a plant expression system and has the processes of glycosylation, disulfide bond formation, post-translational processing of a protein folding stool and the like, the yeast expression system has the advantages of quick growth, easiness in culture, easiness in genetic operation, easiness in screening after transformation and low cost, shorter period and higher efficiency.
The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
Claims (3)
1. A screening device for drought-enduring plants is characterized in that: the simulation box comprises a simulation box (100), wherein a fluorescent lamp (103) is arranged at the top of the simulation box (100), and a sand soil layer (102) is arranged at the bottom of the simulation box; the simulation box (100) is characterized in that the inner cavity of the simulation box (100) is sequentially divided into a plurality of simulation areas (101) along the horizontal direction through a plurality of partition plates, the simulation box (100) is provided with an inclined transparent plate (110) so that a precipitation cavity is formed between the transparent plate (110) and a fluorescent lamp (103), the precipitation cavities in different simulation areas (101) are different in size, and the transparent plate (110) comprises a sealing plate and a precipitation plate provided with through holes; one end of the transparent plate (110) is hinged to the inner wall of the simulation box (100), and the other end of the transparent plate is arranged on the arc-shaped wall of the simulation box (100) in a sliding mode through a rubber sealing piece; the partition plate comprises an upper telescopic plate and a lower telescopic plate, and the transparent plate (110) is arranged between the upper telescopic plate and the lower telescopic plate and is respectively connected with the upper telescopic plate and the lower telescopic plate;
a turbulence component is arranged in the simulation box (100), the turbulence component comprises a rotating shaft (210) transversely arranged in the simulation box (100), the rotating shaft (210) is provided with a horizontal turbulence blade (211), and the horizontal turbulence blade (211) is connected with the rotating shaft (210) through an eccentric hole; the turbulence assembly further comprises a vertical turbulence blade (220), the vertical turbulence blade (220) comprises a shaft body (221) and a connecting sleeve sleeved on the shaft body (221) through an eccentric hole, and the connecting sleeve is provided with a blade; the shaft body (221) is connected with the rotating shaft (210) through an umbrella-shaped gear structure, the umbrella-shaped gear structure comprises a first umbrella-shaped gear arranged on the rotating shaft (210) and a second umbrella-shaped gear arranged on the shaft body (221), and the first umbrella-shaped gear is meshed with the second umbrella-shaped gear; a rotating bearing is arranged on a shaft body (221) of the vertical spoiler blade (220), and the rotating bearing is connected with a supporting piece arranged on the side wall of the partition plate or the simulation box (100);
the screening device for drought-enduring plants further comprises a micro root chamber system (300), wherein the micro root chamber system comprises micro root tubes arranged in a sandy soil layer, a micro CCD camera, a regulating and controlling handle, an image acquisition control device and a computer image processing and analyzing system, and the micro CCD camera, the regulating and controlling handle, the image acquisition control device and the computer image processing and analyzing system are arranged outside the simulation box.
2. A method for screening drought-tolerant plants by using the drought-tolerant plant screening device of claim 1, which comprises the following steps: the method comprises the following steps:
plant sample preparation: selecting a plurality of 2-year-old robust cutting seedlings or seedlings of commonly-used afforestation trees and shrubs, and measuring the initial root conditions and the seedling heights of the seedlings;
desert environment simulation: simulating a desert drought environment in a simulation box (100), wherein the simulation box (100) comprises a plurality of simulation areas (101);
drought tolerance screening: the volumes of the various water reducing cavities are different by adjusting the inclination angle of the transparent plate (110), water is conveyed into the water reducing cavities and the water reducing cavities are filled up, so that different precipitation degrees are set in different simulation areas (101), plants are respectively planted in a sand layer (102) arranged at the bottom of the simulation area (101), under the condition of temperature change of 10-42 ℃, a daylight lamp (103) is used for simulating sunshine for 12-16h/d at the illumination intensity of 4000-; and selecting plants with good root system conditions and good tree growth conditions according to the images obtained by the micro root chamber system (300) processing and analyzing.
3. The method of claim 2, wherein the drought tolerant plant is selected from the group consisting of: the plant sample comprises evergreen arbor: dragon juniper, Japanese pine, Cinnamomum japonicum Thunb, Machilus thunbergii, Zingiber zerumbet Maxim, Ligustrum lucidum ait, evergreen Chinese wax and ilex chinensis Benth; also includes deciduous trees: alder, beech, Sophora japonica, Toona sinensis, Sapindus mukorossi and jujube tree; also includes evergreen shrubs: ramulus et folium Cacaliae Caudatae, Photinia fraseris, flos Rosae chinensis, ramulus Buxi Sinicae, Aleurites fordii, folium seu cortex Nerii, Ligustrum japonicum, and flos Hibisci Mutabilis; also includes deciduous shrubs: malus halliana, Morus alba and Lagerstroemia indica.
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| CN104535752A (en) * | 2014-12-19 | 2015-04-22 | 环境保护部南京环境科学研究所 | Environmental factor-biological interaction simulation device for terrestrial ecological system |
| CN107581040A (en) * | 2017-09-04 | 2018-01-16 | 西北农林科技大学 | The multifarious high flux culture systems of one kind identification Root inheritance and application |
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| CN104535752A (en) * | 2014-12-19 | 2015-04-22 | 环境保护部南京环境科学研究所 | Environmental factor-biological interaction simulation device for terrestrial ecological system |
| CN107581040A (en) * | 2017-09-04 | 2018-01-16 | 西北农林科技大学 | The multifarious high flux culture systems of one kind identification Root inheritance and application |
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| Root and shoot characteristics as selection criteria for drought tolerance in bread wheat (Triticum aestivum L.) at seedling stage under tropical environment;P.K. Kimurto等;《DISCOVERY AND INNOVATION》;20050630;第17卷(第1-2期);78-84 * |
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Inventor after: Liu Zhenxing Inventor after: Wang Lingsheng Inventor after: Zhou Bingtu Inventor after: Huang Qin Inventor after: Yang Xinmou Inventor before: Liu Zhenxing Inventor before: Wang Lingsheng Inventor before: Zhou Bingtu Inventor before: Huang Qin Inventor before: Yang Xingmou |
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Address after: 310016 block B, building 19, Jingfang 6th District, Jianggan District, Hangzhou City, Zhejiang Province Patentee after: Hangzhou Lantian Garden Design and Construction Co.,Ltd. Address before: 310016 block B, building 19, Jingfang 6th District, Jianggan District, Hangzhou City, Zhejiang Province Patentee before: HANGZHOU BLUESKY LANDSCAPE CONSTRUCTION CO.,LTD. |
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