CN112992271A - Method for rapidly predicting crop latitude adaptability indoors - Google Patents

Method for rapidly predicting crop latitude adaptability indoors Download PDF

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CN112992271A
CN112992271A CN202011229587.5A CN202011229587A CN112992271A CN 112992271 A CN112992271 A CN 112992271A CN 202011229587 A CN202011229587 A CN 202011229587A CN 112992271 A CN112992271 A CN 112992271A
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day length
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欧阳鑫昊
黄烯
王向峰
邱蕾蕾
钱崇祯
吴芹芹
王笑颖
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Xiamen University
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Abstract

A method for rapidly predicting latitude adaptability of crops indoors relates to the field of crop genetic breeding. Building a crop day length identification simulator, dividing the crop day length identification simulator into 7 independent growth spaces, and setting different light and dark alternating time for each growth space for simulating 7 kinds of day length at different latitudes; respectively placing the germinated crops into 7 independent growth spaces, collecting and storing crop leaf materials after the crops grow in different illumination lengths, and collecting experimental data of a daily length identification simulator for sorting the crops; identifying the crop day length identification mode of the crop day length identification simulator to obtain the expression of the crop florigen genes at different latitudes; predicting the latitude adaptability of the crops according to the expression of the florigen genes of the crops at different latitudes; and establishing an evaluation system of the prediction accuracy of the crop day length recognition simulator, and quantitatively evaluating the prediction accuracy by using field data. According to the prediction result, two breeding strategies for accelerating the adaptive selection of the crops can be correspondingly provided.

Description

Method for rapidly predicting crop latitude adaptability indoors
Technical Field
The invention relates to the field of crop genetic breeding, in particular to a method for rapidly predicting crop latitude adaptability indoors.
Background
Latitude adaptability means that under the condition of a given latitude, the growth period of a crop variety meets the national variety approval standard of a planting area at the latitude. For example, in the mid-upstream region of the Yangtze river at 30 ° north latitude at a given latitude, the fertility period of indica rice varieties requires up to 150 days. When the growth period of a certain variety is planted in the area and the growth period is about 150 days (the deviation is not more than 5 days), the variety is considered to have better latitude adaptability in the area.
The latitude adaptability of crops is closely related to the yield of crops, and researches show that the length of a growth period is in a positive correlation with the yield of crop varieties (Gao et al, 2014). Therefore, only the variety with better latitude adaptability can ensure the stable yield and the high yield of the crops. Too short growth period can cause crop yield reduction, too long growth period and too low later temperature is not beneficial to crop seed setting.
Under the condition of coping with global warming, the expansion of crop planting areas to high-latitude areas is an effective strategy for ensuring the grain safety. In the future, new agricultural production bases can be developed in high-latitude areas due to the rise of air temperature; crops at mid-low latitudes benefit from increased temperature to extend their planting period. It is expected that how to quickly and efficiently establish a set of selection method for crop latitude adaptability is crucial to ensure food safety in the future expansion process of crop planting areas (Esheld and Lippman, 2019; Sloat et al, 2020).
The length of the growth period of a crop is directly determined by the flowering time of the crop. Many studies have reported that latitude adaptability of crops is closely related to photoperiod genes. In potatoes, allelic mutations of one DOF transcription factor StCDF1 allow a potato to be planted in a wider area than its origin (Kloosterman et al, 2013). In soybean, the J gene is homologous to the arabidopsis thaliana ELF3 gene, and its loss of function prolongs the vegetative growth phase of soybean, increases yield under short sunlight, and promotes expansion of soybean in tropical regions (Lu et al, 2017). Recent studies have shown that, in the domestication of soybeans, mutations in the PRR homologous genes Tof11 and Tof12 allow soybeans to be harvested earlier, while at the same time, improving the adaptability of soybeans at high latitudes (Lu et al, 2020). During early acclimation of maize, the natural variation of the regulatory regions of zmcc 9, zmcc 10 and ZCN8 is of paramount importance for the extended planting of maize from tropical to temperate regions (Yang et al, 2013; Guo et al, 2018). In rice, several studies have reported that Ghd7, Hd1, DTH8(DTH8/Ghd8/LHD1) and DTH7(Ghd7.1/OsPRR37) influence the distribution of rice varieties in geographic locations (Takahashi et al, 2009; Koo et al, 2013; Zhang et al, 2015). Recent studies have also shown that the molecular modules Ghd7-Hd1 and DTH8-Hd1 are very important for modulating the transcription of Ehd1 and Hd3a (Du et al, 2017; Zhang et al, 2017).
Knowledge of geography and astronomy is known: at different latitudes, the length of sunshine (day length) of each day varies periodically in one year, and the different latitudes differ; the higher the latitude, the more drastic the change in the day-to-day of the year. The sunshine length change of the area close to the equator is small and is kept about 12h all year round (figure 1). The sunshine length in high latitude areas changes violently in one year, the summer day length can be more than 16h, the winter day length can be less than 10h, and even in the north and south poles or in the extreme days and the extreme nights; thus, the periodic variation of the day length over the year can be an important indicator of latitude.
Studies have shown that photoperiod genes of different crops are ultimately functional to latitude by directly or indirectly regulating the transcription of florigen gene FT and its homologous genes of different crops (Izawa et al, 2003; Tsuji et al, 2011,2013) (FIG. 2). In plants, photoperiod genes regulate FT or FT homologous genes to sense daily changes in sunshine length (changes in day length). For example, long day plants such as Arabidopsis need to undergo a vernalization process in winter and then receive increasing day length in spring to induce flowering. In arabidopsis, increasing daily growth was accompanied by a gradual induction of the florigen FT gene (Song et al, 2010) (fig. 3). In contrast, typical short-day plants rice grow in the summer, and short-day can promote flowering of rice. The rice recognizes the threshold value of the critical day length through a double-gate control system so as to sense the change of seasons. In japonica rice, Ghd7 and OsGI participate in two optical signal paths for sensing red light and blue light, respectively, to set a threshold for critical day length (threshold 13.5h) (Itoh and Izawa, 2013). When the day is shorter than 13.5h, the transcription of Hd3a of the FT homologous gene can be induced, and flowering is induced (FIG. 4).
Different crops utilize different photoperiod genes to adapt to growth at different latitudes, and even different varieties of the same crop also utilize different photoperiod genes to adapt to different latitudes (Zhang et al, 2015); therefore, there is a need for a method for rapid identification and prediction of latitude adaptability generally applicable to all crops, so as to help breeders modify crop adaptability in response to global warming.
Disclosure of Invention
The invention aims to provide a method for rapidly predicting the latitude adaptability of crops indoors, which can be used for rapidly identifying and predicting the latitude adaptability universally suitable for all crops and thus obtains an optimal adaptive breeding strategy.
The invention comprises the following steps:
1) building a crop day length recognition simulator, which consists of 7 independent growth spaces, wherein each independent growth space is provided with different light and dark alternating time for simulating the length of the day of 7 different latitudes;
2) respectively placing the germinated crops into 7 independent growth spaces of a crop day length recognition simulator, carrying out growth with different illumination lengths, collecting and storing crop leaf materials after the crops grow to the photosensitive period of the crops, and collecting and sorting experimental data of the crop day length recognition simulator;
3) identifying the crop day length identification mode of the crop day length identification simulator to obtain the expression of the crop florigen genes at different latitudes;
4) predicting the latitude adaptability of the crops according to the expression of the florigen genes of the crops at different latitudes;
5) and establishing an evaluation system of the prediction accuracy of the crop day length recognition simulator, and quantitatively evaluating the prediction accuracy by using field data.
In step 1), the independent growth space can be an independent artificial climate chamber, an independent illumination incubator or an independent space with at least independent control of illumination length; the specific method for building the crop day length recognition simulator can be as follows: 24h is set as a periodic light-dark alternating environment in each independent growth space, and the light-dark alternating time of 7 independent growth spaces is respectively set as 10h light/14 h dark, 12h light/12 h dark, 13h light/11 h dark, 13.5h light/10.5 h dark, 14h light/10 h dark, 14.5h light/9.5 h dark and 16h light/8 h dark; the illumination intensity is more than 300 mu mol m-2s-1。
In step 2), after the crop grows to the photosensitive period of the crop, collecting crop leaf material and storing, preferably: the rice can grow for 33 days, and after the rice is irradiated for 3 hours on day 34 (ZT3), leaf materials are collected and stored in liquid nitrogen; placing the corns in a greenhouse with different illumination lengths until the corns grow to 8 leaves (about 40 days), and sampling at ZT 1; soybeans were grown in a greenhouse with different illumination lengths for 14 days and sampled at ZT 4.
In step 3), the expression of the obtained plant florigen genes at different latitudes can be detected by a fluorescent quantitative PCR experiment to detect the expression condition of the plant florigen genes, a curve is drawn, and the plants are divided into 3 types according to a day length identification mode: 1. critical day length can be identified; 2. the gradual change day length can be identified; 3. insensitivity to day length and continuous high expression;
the specific conditions for classifying the crops into 3 types according to the day length identification mode can be as follows:
(1) when the difference of the florigen gene expression of two adjacent treatments is more than ten times, the crop is considered to be capable of identifying the critical day length, and the threshold value of the critical day length is the activated gene expression of the two adjacent treatments;
(2) when the anthocyanin expression gradually changes with the change in day length and the difference between adjacent treatments is less than 10-fold, the crop is considered to recognize the gradual day length;
(3) when the expression of anthocyanin hardly changes with the change of day length and is always at high expression, the crop is considered to be insensitive to the change of day length.
In step 4), the specific method for predicting the florigen gene expression at different latitudes may be: expression of florigen gene according to 7 different day length treatments obtained in step 3), whereas day length change in one year is known at any latitude; selecting three latitudes of high (47 degrees north latitude), medium (35 degrees north latitude) and low (23 degrees north latitude), and expressing the daily lengths of the three latitudes from 3 months to 12 months by a curve to obtain field experimental data; meanwhile, the florigen genes in each crop are divided into three levels of high expression, low expression and no expression, the expression of the florigen genes of different species is divided into the 3 levels by combining the day length change of each latitude, the colors of different depths are used for representing under the corresponding day length, and then drawing graphs are compared, so that the latitude adaptability result of the crop can be obtained.
In step 5), the specific method for establishing the evaluation system for the prediction accuracy of the crop day length recognition simulator may be: collecting materials from the seedling stage to the flowering stage by using crops growing in the field at a given latitude, and detecting the expression of florigen genes; the day length situation of the current day of material drawing can be known through the material drawing time and the latitude, and approximate estimation data is obtained by using the expression approximate estimation of florigen genes corresponding to the day length in the experimental data of the crop day length identification simulator; then carrying out normalization processing on the field experimental data and the maximum value of the approximate estimation data; the other data are converted by multiplying by a conversion coefficient c (c 1/max), and then a determination coefficient R is calculated using a linear regression model in R software2
Figure BDA0002764723730000041
And p value to evaluate the accuracy of the DEAS prediction (SST ═ sum of squares; SSR ═ sum of squares due to regression).
Based on the method for rapidly predicting the latitude adaptability of the crop indoors, the invention also provides a breeding strategy for accelerating the adaptive selection of the crop, and the breeding strategy can be divided into the following two types:
a: for crops (rice) with a large number of cloned photoperiod genes, molecular design breeding can be carried out by using a CRISPR/Cas9 gene editing technology; based on the method for rapidly predicting the latitude adaptability of the crop indoors, the steps 2) and 3) can identify the day length recognition mode of the reference variety at the given latitude, the photoperiod genotype of the reference variety is known through high-throughput sequencing, the photoperiod genotype of the crop material is further known through the high-throughput sequencing, the crop material is modified through a CRISPR/Cas9 gene editing technology according to the genotype of each gene of the reference variety, and finally, the crop material consistent with the day length perception mode of the reference variety is obtained;
b: for new crops with functional photoperiod genes not cloned yet, reference varieties from different latitudes can be analyzed through the steps 2) and 3) of the method for rapidly predicting the latitude adaptability of the crops indoors, day length perception modes with adaptability at the latitudes are represented, breeding materials and genetic groups are classified according to the day length recognition mode of the reference varieties, and according to the mode, resource libraries can be established for the breeding intermediate materials and the genetic groups according to different latitudes; through the strategy, a breeding resource library with determined latitude adaptability can be generated, and guarantee is provided for accelerating crop variety breeding under global climate change.
The invention establishes the relation among the daily growth identification mode of the crop, the dynamic transcription and the latitude of the florigen gene by analyzing the mechanism of the crop identification daily growth, predicts the latitude adaptability of the crop by establishing a crop daily growth identification simulator, and can correspondingly provide two breeding strategies for accelerating the selection of the crop adaptability according to the prediction result. Compared with the prior art, the method can quickly predict the latitude adaptability of the crops by utilizing the crop materials about 33 days indoors, and accelerate the adaptability selection of the crops in breeding. The method is simple, has wide applicability, can be generally suitable for quickly identifying and predicting the latitude adaptability of all crops, and has an important effect on improving the stable yield and the high yield of the crops.
Drawings
Fig. 1 is a graph showing the variation of sunshine length in one year at different latitudes.
Fig. 2 is a photoperiod network of different crops. The boxes in the figure represent the florigen genes of different crops, so it can be seen that the photoperiod network nodes in different plants are all florigen genes.
FIG. 3 is a schematic representation of the induction of florigen gene expression by Arabidopsis thaliana with increasing sensory day length.
FIG. 4 is a schematic representation of rice induction of florigen gene expression by sensing critical day length.
Fig. 5 is a schematic diagram of the architecture of a crop day length identification simulator. Wherein the horizontal line background represents illumination, the vertical line background represents darkness, and seven independent growth spaces respectively simulate 24h of seven different sunlight length changes.
FIG. 6 is a diagram of three day length recognition modes identified by experimental data collection and analysis of a crop day length recognition simulator.
FIG. 7 is experimental data based on a day length recognition simulatorAnd converting into a latitude adaptability schematic diagram of the crop. The oryza sativa florigen gene 10 > Hd3a/UBQ (x 10) in FIG. 6-4) Defined as no expression, 100 > Hd3a/UBQ (x 10)-4) Under expression, Hd3a/UBQ (> 10) (x 10)-4) High expression is defined as > 100; the maize florigen gene 100 > ZCN8/UBQ (x 10)-5) Defined as no expression, 1000 > ZCN8/UBQ (x 10)-5) > 100 is defined as low expression, ZCN8/UBQ (x 10)-5) High expression is defined as > 1000; soybean florigen gene GmFT2a/Tubulin (x 10)-4) High expression is defined as > 1000.
FIG. 8 is a schematic diagram of a crop day length identification simulator based prediction of latitude fitness of a crop.
FIG. 9 is a graph of an evaluation system for establishing the prediction accuracy of a crop day length recognition simulator using field data. Rice planted in Wenjiangjiang (N30 degrees 40 ', E103 degrees 50') of different genotypes, sampling from the seedling stage to the heading stage, expressing the florigen gene Hd3a, and normalizing the maximum value, as shown by the open dots in the figure; the maximum value is normalized according to the day length of the region at different times and the Hd3a expression obtained by the day length recognition simulator, as shown by a broken line in the figure.
FIG. 10 is a schematic diagram of a breeding strategy for establishing adaptive selection of crops using a crop day length recognition simulator based on crops (rice) in which a large number of photoperiod genes have been cloned. The different line directions represent different latitudes or ecological planting areas, and the crop day length recognition simulator can be used for investigating the day length recognition mode of the reference material and the genotype of the photoperiod gene, so that the latitude adaptability modification of the crop can be carried out by combining the CRISPR/Cas9 technology.
FIG. 11 is a schematic diagram of a breeding strategy for establishing crop adaptive selection using a crop day length recognition simulator based on new crops for which functional photoperiod genes have not been cloned. Wherein panel a represents breeding material and populations; b, different line directions of the graph represent different latitudes or ecological planting areas, and a crop day length identification simulator is used for investigating a day length identification mode of a reference material; and the graph C is used for carrying out latitude adaptability classification on the breeding materials and the population according to the day length identification mode of the reference materials, and providing germplasm resources with different latitude adaptability for subsequent breeding.
FIG. 12 is a flow chart of a method for predicting crop latitude fitness based on a crop day length identification simulator.
FIG. 13 is a flow diagram of a breeding strategy for accelerating crop adaptability selection based on a crop day length recognition simulator.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to the accompanying drawings.
The embodiment of the invention comprises the following steps:
(1) and (3) constructing a facility of the crop day length identification simulator: first, 7 independent climatic chambers or light incubators, or at least independent spaces with independently controlled light length, need to be established for crop seedling growth (fig. 5). Each independent space is provided with an environment with 24h as periodic light and darkness alternation, and the light lengths of 7 independent growth spaces are respectively set to 10h (10h light/14 h darkness), 12h (12h light/12 h darkness), 13h (13h light/11 h darkness), 13.5h (13.5h light/10.5 h darkness), 14h (14h light/10 h darkness), 14.5h (14.5h light/9.5 h darkness) and 16h (16h light/8 h darkness). The illumination intensity is more than 300 mu mol m-2s-1. Thus, the entire crop day length identification simulator consists of 7 independent growth spaces (fig. 5). 7 different sunshine lengths were simulated, respectively.
(2) Collecting experimental data of a crop day length identification simulator: respectively placing the germinated crops into 7 independent growth spaces of a crop day length recognition simulator, carrying out growth with different illumination lengths, collecting and storing crop leaf materials after the crops grow to the photosensitive period of the crops, and collecting and sorting experimental data of the crop day length recognition simulator; among them, preferred are: the rice can grow for 33 days, and after 3 hours of illumination on the 34 th day (ZT3), leaf materials are collected and stored in liquid nitrogen; the corn can be placed in a greenhouse with different illumination lengths to be grown to 8 leaves (about 40 days), and the corn is sampled at ZT 1; soybeans were grown in a greenhouse with different illumination lengths for 14 days and sampled at ZT 4. And detecting the florigen gene expression condition of the crops through a fluorescent quantitative PCR experiment.
(3) The crop day length identification simulator identifies the crop day length identification mode: the experimental data collected by the crop day length identification simulator are collated to obtain a result as shown in fig. 6, when the difference of florigen gene expression of two adjacent treatments is more than ten times, the crop is considered to be capable of identifying the critical day length, and the threshold value of the critical day length is the activated gene expression of the two adjacent treatments (fig. 6A-B). When florigen expression gradually changed with the change in day length, and the difference between adjacent treatments was less than 10-fold, it was considered that the crop could recognize the gradual day length (fig. 6D-E). When the florigen expression hardly changed with the change in the day length and was always at high expression, the crop was considered insensitive to the change in the day length (FIGS. 6G-H). Therefore, the built day length identification simulator can roughly divide the day length identification modes of crops into three types: 1: identifying a critical day length (fig. 6C); 2: identifying a fade day length (fig. 6F); 3: insensitivity to day length continued to be highly expressed (FIG. 6I).
(4) The crop day length identification simulator predicts the latitude adaptability of crops: the experimental data shown in fig. 6 are collected by different crop seedlings through a crop day length recognition simulator, a set of methods for predicting the expression of the florigen genes at different latitudes is invented based on the experimental data, and finally, the latitude adaptability of crops can be predicted according to the expression modes of the florigen genes at different latitudes. The method for predicting the expression of florigen genes at different latitudes is as follows: from FIG. 6, it can be seen that the florigen gene expression of any crop was observed at 7 different day length treatments, and the change in day length in one year was known at any latitude (FIG. 7). Therefore, three latitudes, i.e., high (47 ° north latitude), medium (35 ° north latitude), and low (23 ° north latitude), are selected, and the daily lengths of these three latitudes from 3 months to 12 months are represented by a curve (fig. 7). Meanwhile, according to the results in fig. 6, the florigen genes in each species were classified into three grades of high expression, low expression, and no expression. Combining FIG. 6 and the variation of the day length at each latitude, the expression of florigen genes of different species was classified into the above 3 classes and expressed with colors of different depths at the corresponding day lengths, and the results shown in FIG. 7 were obtained.
For example, after 19 days 8.8 in FIG. 8, the day length of N47 ° gradually decreased from 14h to 13.5h on 29 days 8.8, so that the expression of the florigen gene of a given crop at this latitude during this period can be approximated by the expression of the florigen gene of 13.5h of the same crop in FIG. 6. Similarly, from day 17 of 3 months to day 4 of 4 months, the day length of N47 ° increased from 12h to 13h, and the expression of the florigen gene at 13h in FIG. 6 was approximated. When the expression data of fig. 6 is divided into levels and represented in fig. 7 and 8 with different depths of color, the prediction data of fig. 7 and 8 is equivalent to a heat map of florigen gene expression as a function of day length.
(5) Establishing an evaluation system of the prediction accuracy of the crop day length recognition simulator: using crops grown in the field at a given latitude, material was harvested from the beginning of the seedling stage until flowering and the expression of the florigen gene was examined. The day length of the day of material drawing can be known through the time and latitude of material drawing, for example, the first sampling point in fig. 9 is at 40 ° N30', and the sampling is performed at 9 th 6, and the day length of the day is 14.1h through a day length calculation formula, and the expression of the florigen gene at 14h in fig. 6 can be used for approximate estimation in the change process of 14.5-14 h. Further, the maximum values of the field experimental data and the approximate estimation data are normalized. The other data is converted by multiplying by a conversion coefficient c (c 1/max). Next, using a linear regression model in the R software, the determination coefficient R is calculated2
Figure BDA0002764723730000071
And p value to evaluate DEAS prediction accuracy (SST ═ sum of squares; SSR ═ sum of squares due to regression) (fig. 9).
(6) Accelerating the adaptive selection of crops by utilizing a crop day length recognition simulator: (4) predicting the latitude adaptability of the crop is based on the crop day length identification modes identified in (2) and (3). Therefore, two strategies are generalized to accelerate the adaptive selection of crops by using the day length identification mode of the crops:
a: for crops (rice) with a large number of cloned photoperiod genes, molecular design breeding can be carried out by using CRISPR/Cas9 gene editing technology. Based on (2) and (3), the day-length identification mode of the reference variety at a given latitude can be identified, and the photoperiod genotype of the reference variety is known through high-throughput sequencing. Further using high throughput sequencing to learn the photoperiod genotype of the crop material. Further, according to the genotypes of the genes of the reference varieties, the crop material is modified by using a CRISPR/Cas9 gene editing technology, and finally, the crop material which is consistent with the day-length perception mode of the reference varieties is obtained (figure 10).
B: for new crops in which functional photoperiod genes have not been cloned, reference varieties from different latitudes can be analyzed through (2) and (3), and adaptive day-length perception modes at the latitudes can be characterized. Furthermore, the breeding materials and the genetic groups are classified according to the day length identification mode of the reference variety, and according to the mode, resource libraries can be established for the breeding intermediate materials and the genetic groups according to different latitudes. Through the strategy, a breeding resource library with determined latitude adaptability can be generated, and guarantee is provided for accelerating crop variety breeding under global climate change (figure 11).
In summary, the invention develops two sets of technical schemes with different targets by utilizing a crop day length identification simulator, a method for identifying a corresponding derived crop day length identification mode, a method for predicting crop latitude adaptability and the like.
A: a method for predicting crop latitude adaptability based on a crop day length recognition simulator (fig. 12): through the steps of utilizing the crop day length identification simulator (step 1), collecting the experimental data of the crop day length identification simulator (step 2), and further through a crop latitude adaptability prediction method (step 4), the method can utilize the expression patterns of florigen genes at different latitudes to represent the latitude adaptability of crops (step 4). Meanwhile, a set of statistical method for quantitatively evaluating and predicting accuracy by using field data is invented (step 5).
B: breeding strategies based on crop day length recognition simulator accelerated crop adaptability selection (fig. 13): by utilizing a crop day length identification simulator (step 1), collecting experimental data of adaptive reference materials with different latitudes (step 2), further utilizing (step 3) a day length identification mode for identifying the reference materials, and finally accelerating crop adaptive selective breeding according to the strategy of the step (step 6).
The following gives a flow of a specific example.
Scheme 1: obtaining of test materials
Rice materials used in FIGS. 6 and 9 of the present invention, Hd1DTH8Ghd7, Hd1DTH8Ghd7, Hd1DTH8Ghd7, Hd1DTH8Ghd7, and NILHd1dth8Ghd7Corresponding to the materials Hd1DTH8, Hd1DTH8, Hd1DTH8, Hd1DTH8 and NIL in the previous publications, respectivelydH2-1(Du et al, 2017). Hd1DTH8Ghd7(Hd1DTH8) is a wild-type material (genetic background: Dongjin), and Hd1DTH8Ghd7(Hd1DTH8) is a double-mutant material of Hd1 and DTH8 in the Dongjin background. The genotype of the photoperiod flowering gene for these materials is known.
The maize material T32 and LX9801 used in FIG. 6 of the present invention are inbred lines in maize breeding. T32 is a hot strip material suitable for tropical planting at low latitudes, and flowers particularly late in middle and high latitudes, so the hot strip material does not have adaptability at the middle and high latitudes, and LX9801 is a temperate strip material, has moderate flowering time in the middle and high latitudes and early flowering time in the low latitudes, and can be used as an early maturing variety. The genotype of the photoperiod flowering gene for these materials was unknown.
The soybean variety Heilongjiang 64 used in the invention in figure 6 is temperate material, has moderate flowering time in high latitude areas, has early flowering time in middle and low latitude areas, and can be used as an early-maturing variety. The genotype of the photoperiod flowering gene for these materials was unknown.
And (2) a flow scheme: collecting experimental materials in a crop day length recognition simulator
Mixing rice material (Hd1DTH8Ghd7, Hd1DTH8Ghd7, NIL)Hd1dth8Ghd7) 6 total materials of corn (T32, LX9801) and soybean (Heilongjiang 64) were placed in a growth chamber (FIG. 5) for 10h, 12h, 13h, 13.5h, 14h, 14.5h, and 16h illumination time for the corresponding days, and samples were taken at the corresponding times (2).
And (3) a flow path: as can be seen from FIG. 2, the photoperiod signals of almost all plants eventually converge on the florigen gene. The florigen genes of the current model plant Arabidopsis thaliana and important crops have been cloned (Yang et al, 2014; Song et al, 2015; Lu et al, 2020). Their names and gene IDs are as follows:
growth of ArabidopsisThe gene of the flower essence: FT (AT1G65480)
Florigen gene of rice: hd3a (Os06g0157700)，RFT1(Os06g0157500)
Florigen gene of maize: ZCN8(GRMZM2G179264)，ZCN12(GRMZM2G103666)
Floral-forming gene of soybean: GmFT2a (Glyma.16G150700)
Sorghum florigen gene: SbCN8(Sb09g025760)，SbCN12(Sb03g034580)
Florigen gene of wheat: FT1(DQ890162)
And (4) a flow chart: further extracting RNA from the material collected in scheme 2 using a Promega RNA extraction kit using oligo (dT) (Promega) GoScriptTMReverse transcription kit cDNA was synthesized by reverse transcription from 800ng total RNA. qPCR was performed using TaqMan probe method to detect the expression of rice Hd3a relative to UBQ (FIGS. 6A, D and H); qPCR was performed using the SYBR dye method to detect the expression of maize ZCN8 relative to UBQ and the expression of soybean GmFT2a relative to Tubulin (fig. 6B, E and G).
And (5) a flow chart: by using the data (fig. 6) obtained in the process 4, according to the method for predicting the latitude adaptability of the crop by the crop day length recognition simulator, under the selected three latitudes of high, medium and low, the expression data of fig. 6 is expressed under different day lengths of different latitudes, so as to obtain the result shown in fig. 7, and the method for predicting the latitude adaptability of the crop by the day length recognition simulator is established by combining the data of fig. 6 and the data of fig. 7, as shown in fig. 8.
And (6) a flow path: in order to verify the accuracy of predicting the florigen gene expression of the field rice material by the day-length recognition simulator, 4 materials with different genotypes are planted in Wenjiang river, the materials are sampled from the seedling stage to the heading stage, then RNA of the rice at each stage is extracted, reverse transcription is carried out, the expression of the florigen gene Hd3a relative to a reference gene UBQ is detected, the maximum value is regarded as 1, and other data are multiplied by a conversion coefficient to obtain corresponding data, as shown by the open dots in figure 9; the expression data of Hd3a is obtained from the day length identification simulator, the corresponding expression data of Hd3a in the day length identification simulator is taken according to the day length of the sampling day, the maximum value is also taken as 1, and the other data are multiplied by the conversion coefficient to obtain the corresponding data, as shown in the broken line diagram of FIG. 9.

Claims (10)

1. A method for rapidly predicting the latitude adaptability of crops indoors is characterized by comprising the following steps:
1) building a crop day length recognition simulator, which consists of 7 independent growth spaces, wherein each independent growth space is provided with different light and dark alternating time for simulating the length of the day of 7 different latitudes;
2) respectively placing the germinated crops into 7 independent growth spaces of a crop day length recognition simulator, carrying out growth with different illumination lengths, collecting and storing crop leaf materials, and collecting and arranging experimental data of the crop day length recognition simulator;
3) identifying the crop day length identification mode of the crop day length identification simulator to obtain the expression of the crop florigen genes at different latitudes;
4) predicting the latitude adaptability of the crops according to the expression of the florigen genes of the crops at different latitudes;
5) and establishing an evaluation system of the prediction accuracy of the crop day length recognition simulator, and quantitatively evaluating the prediction accuracy by using field data.
2. The method for fast indoor crop latitude adaptability prediction according to claim 1, characterized in that in step 1), the independent growth space adopts an independent artificial climate chamber, an independent light incubator or an independent space with at least independent control of light length.
3. The method for rapidly predicting the latitude adaptability of the crop indoors according to claim 1, wherein in the step 1), the specific method for constructing the crop day length recognition simulator is as follows: 24h is set as a periodic light-dark alternating environment in each independent growth space, and the light-dark alternating time of 7 independent growth spaces is respectively set as 10h light/14 h dark, 12h light/12 h dark, 13h light/11 h dark, 13.5h light/10.5 h dark, 14h light/10 h dark, 14.5h light/9.5 h dark and 16h light/8 h dark; high illumination intensityAt 300. mu. mol m-2s-1。
4. The method for rapidly predicting the latitude adaptability of the crop indoors as claimed in claim 1, wherein in the step 1), in the step 2), the collecting crop leaf material is collected after the crop leaf material is grown to a sensitive period of the crop.
5. The method for rapidly predicting the latitude adaptability of the crop indoors according to claim 1, wherein in step 1), in step 2), the preservation adopts liquid nitrogen preservation.
6. The method for rapidly predicting the latitude adaptability of the crops indoors as claimed in claim 1, wherein in the step 3), the expression of the florigen genes of the crops at different latitudes is obtained by detecting the florigen gene expression condition of the crops through a fluorescent quantitative PCR experiment, drawing a curve, and dividing the crops into 3 types according to a day length recognition mode: 1. critical day length can be identified; 2. the gradual change day length can be identified; 3. insensitivity to day length persists with high expression.
7. The method for rapidly predicting the latitude adaptability of the crop indoors according to claim 1, wherein in the step 3), the concrete conditions for classifying the crop into 3 types according to the day length identification mode are as follows:
(1) when the difference of the florigen gene expression of two adjacent treatments is more than ten times, the crop is considered to be capable of identifying the critical day length, and the threshold value of the critical day length is the activated gene expression of the two adjacent treatments;
(2) when the anthocyanin expression gradually changes with the change in day length and the difference between adjacent treatments is less than 10-fold, the crop is considered to recognize the gradual day length;
(3) when the expression of anthocyanin hardly changes with the change of day length and is always at high expression, the crop is considered to be insensitive to the change of day length.
8. The method for rapidly predicting the latitude adaptability of the crop indoors according to claim 1, wherein in the step 4), the specific method for predicting the expression of the florigen gene at different latitudes is as follows: expression of florigen gene according to 7 different day length treatments obtained in step 3), whereas day length change in one year is known at any latitude; and simultaneously, dividing the florigen genes in each crop into three grades of high expression, low expression and no expression, dividing the expression of the florigen genes of different species into the 3 grades by combining the change of the day length of each latitude, expressing the three grades by colors of different depths under the corresponding day length, and comparing drawn graphs to obtain the latitude adaptability result of the crop.
9. The method for rapidly predicting the latitude adaptability of the crop indoors according to claim 1, wherein in the step 5), the specific method for establishing the evaluation system of the prediction accuracy of the crop day length recognition simulator is as follows: collecting materials from the seedling stage to the flowering stage by using crops growing in the field at a given latitude, and detecting the expression of florigen genes; the day length situation of the current day of material drawing can be known through the material drawing time and the latitude, and approximate estimation data is obtained by using the expression approximate estimation of florigen genes corresponding to the day length in the experimental data of the crop day length identification simulator; then carrying out normalization processing on the field experimental data and the maximum value of the approximate estimation data; the other data were converted by multiplying by a conversion coefficient c (c 1/max), and then using a linear regression model in R software, a determination coefficient was calculated
Figure FDA0002764723720000021
And p value to evaluate the accuracy of the DEAS prediction (SST ═ sum of squares; SSR ═ sum of squares due to regression).
10. A breeding strategy for accelerating the adaptive selection of crops, which is characterized by the following two categories:
a: for crops with a large number of cloned photoperiod genes, molecular design breeding is carried out by using a CRISPR/Cas9 gene editing technology; identifying the day length recognition mode of a reference variety at a given latitude based on steps 2) and 3) of the method according to claim 1, obtaining the photoperiod genotype of the reference variety through high-throughput sequencing, further obtaining the photoperiod genotype of the crop material through the high-throughput sequencing, modifying the crop material by using a CRISPR/Cas9 gene editing technology according to the genotypes of all genes of the reference variety, and finally obtaining the crop material consistent with the day length perception mode of the reference variety;
b: for new crops for which functional photoperiod genes have not been cloned, reference varieties from different latitudes are analyzed through steps 2) and 3) of the method according to claim 1, day length perception modes with adaptability at the latitudes are characterized, breeding materials and genetic groups are classified according to the day length recognition mode of the reference varieties, and according to the mode, resource libraries are established for the breeding intermediate materials and the genetic groups according to different latitudes; through the strategy, a breeding resource library with determined latitude adaptability is generated, and guarantee is provided for accelerating crop variety breeding under global climate change.
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