CN113692966A - Distant hybridization method for rice and sorghum - Google Patents
Distant hybridization method for rice and sorghum Download PDFInfo
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Abstract
The invention relates to a distant hybridization method for rice and sorghum, belonging to the field of distant hybridization breeding of rice, wherein the distant hybridization of rice and sorghum becomes one of effective ways for solving the problems of narrow genetic basis of cultivated rice, expanding genetic variation and creating new germplasm resources, but the distant hybridization and outcrossing seed setting rate of rice and sorghum is very low, and the reasons are mainly that the conventional castration method for rice has large damage to pistil stigma and the sorghum pollen collection time and method are incorrect. Aiming at the problems, the invention adopts an obliquely upward 45-degree glume shearing method, after glume shearing of each ear is finished, a 600-fold liquid of 3% gibberellic acid emulsifiable solution is uniformly sprayed at a point, residual stamen anthers ejecting glumes are removed by pinching or finger flicking before 7 o 'clock in the next morning, bloomed sorghum ears are taken down at 6 o' clock every day and are put into a foam box with wet tissue laid at the bottom, and pollination is carried out at 30 minutes or so at 8 am.
Description
Technical Field
The invention relates to a distant hybridization method for rice and sorghum, belonging to the field of rice hybridization breeding.
Background
Rice (Oryza sativa L.) as a ration for 50% of the population all over the world has the characteristics of wide planting area, high yield and abundant germplasm genetic diversity. The rice is originated in China, the total yield of the rice is the first world, the advantages of hybrid rice are technically reputed to the world, but with the long-term large-area planting of the cultivated rice, the genetic difference among varieties becomes small, the yield increasing potential is slow, the resistance to plant diseases and insect pests is weakened, the continuous healthy and stable development of the national food strategy is not facilitated, and the national food safety is influenced.
Three stages of rice development, namely utilization of inter-breed, inter-subspecies and distant heterosis, were proposed by father Yuanyin Hopugilistica of Chinese hybrid rice in 1978. The utilization research of the hybrid vigor between varieties and subspecies is always in the top of the world in China, but the germplasm innovation between varieties and subspecies is difficult to make a major breakthrough at present, so that the homogenization phenomenon of the current rice variety breeding is increasingly serious. The distant hybridization of rice will become one of the effective ways to solve the narrow hereditary basis of the cultivated rice, expand the hereditary variation and create new germplasm resources.
Sorghum (Sorghum bicolor L.) C4 crop has many excellent properties of high photosynthetic efficiency, smooth operation of photosynthetic products, good bearing property, good plant leaf shape, thick and strong stems, developed root system, strong lodging resistance, fertilizer and drought resistance, waterlogging resistance, disease resistance, strong saline-alkali resistance and the like, and the distant hybridization of the rice Sorghum can solve the adverse effects of narrow genetic basis of cultivated rice, poor germplasm resources, small genetic difference among different varieties of interspecific hybridization of the cultivated rice, slow yield increase potential and the like, and especially has huge potential and application prospect in the aspects of stress resistance and ultrahigh yield cultivation of the rice.
However, genetic difference between sorghum and rice is large, the maturing rate of hybridization between sorghum and rice is very low under the condition that the rice is not damaged and emasculated, while the traditional methods of traditional warm soup emasculation, manual emasculation, conventional glume cutting and water spraying finger emasculation and the like are adopted, so that the rice pistil is damaged in different degrees, the pollination channel is not smooth and the like, the outcrossing maturing rate is further reduced, a large number of tests show that rice sorghum distant hybrid progeny cannot be obtained basically by adopting the warm soup emasculation method, and meanwhile, the time and the method for collecting sorghum pollen have great influence on the outcrossing maturing rate between sorghum and rice.
Disclosure of Invention
The invention aims to: aiming at the problem of low outcrossing bearing rate of distant hybridization of rice and sorghum, the distant hybridization method of rice and sorghum is provided, the method comprises the steps of picking up small ears of target rice ears in the afternoon of the previous day, selecting 20-25 small ears capable of flowering the next day, removing other small ears of the same ears, shearing the retained small ears along the middle part 1/2 by adopting an obliquely upward 45-degree angle glume shearing method, adopting 600 times of 3% gibberellic acid emulsifiable solution to carry out uniform point spraying treatment on all the small ears after the glume shearing of each ear, hanging and marking, pinching or finger flicking the residual stamens extending out of glumes before 7 o's in the next morning and sleeving a paper bag, when the sorghum blooms at the early morning of 6, cutting down flowering sorghum ears, putting the flowering ears into a foam box with a bottom spreading towel, covering the foam box, lightly conveying the cotton ears to the rice field at 8 o's and 30 th o's in the morning, and pollinating the cowhide, gently taking down the sorghum stalks which are scattered with a large amount of pollen, inserting the sorghum stalks into a kraft paper bag, and quickly rotating to achieve the purpose of uniformly scattering the pollen, wherein the pollination is repeatedly carried out for a plurality of times in order to ensure the pollination strength. The method can obviously improve the outcrossing maturing rate of the rice and the sorghum, greatly reduce the labor intensity of operators and greatly improve the castration work efficiency of the rice.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
in order to increase the probability of the gibberellic acid liquid medicine infiltrating to the filaments of the rice stamens and increase the area of a single glume pollination channel, a glume shearing method of 45 degrees upwards is adopted for each spikelet along the middle part of the spike, so that on one hand, the liquid medicine can be ensured to infiltrate to the filaments of the rice stamens, and on the other hand, the glume pollination channel area can be maximized, thereby greatly improving the success probability of cross pollination.
In order to reduce the damage of warm water castration, manual castration and water spraying finger stampings to rice pistils, gibberellic acid is utilized to accelerate cell elongation, stamen filaments can be rapidly stretched, residual anthers of the stamens can be ejected out of spikelet glumes by the stretched filaments, a large number of experiments show that the dilution concentration of 3% gibberellic acid missible oil is 600 times that of the solution, the residual anthers can be rapidly removed by pinching or finger flicking before pollen is scattered 7 days later, castration can basically achieve nondestructive operation on the pistils, the castration efficiency of the rice can be greatly improved, meanwhile, the pistil stigma is further infiltrated into low-concentration gibberellic acid liquid medicine to remarkably improve the stigma viability, and therefore, the rice sorghum distant hybridization heterozygosis maturing rate is improved.
In order to improve the vitality of sorghum pollen and ensure the pollination strength, the invention determines that the sorghum pollen is taken at about 6 am, the sorghum blooms but the stamens do not shed a large amount of pollen, the loss of the pollen of the ear is small, the taken sorghum ear is put into a bottom wet paper towel spreading foam box to simulate a high-humidity environment suitable for sorghum pollen shedding, the sufficient amount of the pollen of the sorghum ear taken at about 30 minutes at 8 am is ensured, the high pollen amount can be ensured by pollination, the probability that the sorghum pollen falls into the stigma of rice is increased, and repeated pollination is adopted for improving the distant hybridization success rate of the rice sorghum.
Detailed Description
Example 1
Influence of different emasculation modes on rice and sorghum hybrid outcrossing setting rate
The test method comprises the following steps: selecting a rice variety as newly selected japonica 16-287, selecting a sorghum variety as local dwarf glutinous sorghum soil, and adopting emasculation modes as follows: the artificial emasculation, the warm soup emasculation (cutting glumes after scalding for 5 minutes at the water temperature of 45 ℃), the conventional glume cutting and water spraying finger emasculation, and the glume cutting at an angle of 45 degrees obliquely upwards by using a liquid point spraying emasculation method of 3% gibberellic acid emulsifiable solution 600 times.
TABLE 1 influence of different detasseling patterns on outcrossing setting percentage
| Castration method | Outcrossing maturing rate (%) |
| Manual castration | 0.53b |
| Removing stamen from warm soup | 0.00c |
| Finger elastic male part for conventional glume cutting and water spraying | 0.13c |
| 45-degree oblique glume shearing gibberellic acid emasculation | 1.07a |
As can be seen from Table 1, different emasculation modes have obvious influence on the outcrossing maturing rate of the rice sorghum, the outcrossing maturing rate of the outcrossing by the glume-cutting 3% gibberellic acid missible oil of which the angle is 45 degrees in the oblique direction is up to 1.07% in 600-time liquid point spraying treatment, the difference is obvious compared with other treatment, secondly, the artificial castration and the conventional glume cutting and water spraying finger castration treatment are carried out, the difference between the outcrossing seed setting rate of 0.53 percent and the outcrossing seed setting rate of 0.13 percent is obvious, while the male removal treatment by warm soup can not form hybrid progeny, the outcrossing setting rate is 0, tests show that the rice sorghum distant hybridization can not adopt the warm soup male removal method, the analysis reasons are mainly caused by pistil damage and pollen channel obstruction, the manual castration and glume-shearing water-spraying finger castration method has the situation that pistils are damaged to different degrees, but the pistil damage is the least by adopting the 600-time liquid spraying castration method of glume-shearing 3% gibberellic acid emulsifiable solution at an oblique 45-degree angle, and meanwhile, a smooth pollination channel can be maintained, so the outcrossing maturing rate is high.
Example 2
Comparative test of emasculation efficiency of different emasculation modes
The test method comprises the following steps: selecting a rice variety as newly selected japonica 16-287, selecting a sorghum variety as local dwarf glutinous sorghum soil, and adopting emasculation modes as follows: the artificial emasculation, the hot water emasculation, the conventional glume cutting and water spraying finger emasculation, and the conventional glume cutting and water spraying finger emasculation method with 600-time liquid point spraying of the 3% gibberellic acid emulsifiable solution, wherein the same pollination mode is adopted, each treatment is repeated for 3 times, each emasculation is repeated for 10 ears, each ear retains 25 small ears, the operation time of each treatment is accurately timed, the operation time is accurate to the second, and the emasculation efficiency of each treatment is compared.
TABLE 2 comparison of castration efficiency between different castration modes
| Castration method | Emasculation efficiency (ear/second) |
| Manual castration | 328.7b |
| Removing stamen from warm soup | 390.7a |
| Finger elastic male part for conventional glume cutting and water spraying | 282.7c |
| 45-degree oblique glume shearing gibberellic acid emasculation | 88.7d |
From table 2, it can be known that the working efficiency of different emasculation modes is very different, the emasculation time of a single spike is shortest when the single spike is processed by adopting the 600-time liquid-point spraying emasculation method of the 3% gibberellic acid emulsifiable solution of the 45-degree angle chaff in the oblique direction, the time consumption of each spike is only 88.7 seconds, the efficiency is improved by 3.7 times compared with the efficiency of artificial emasculation processing, and is improved by 3.19-4.41 times compared with other processing efficiency, and test results show that the emasculation working efficiency can be greatly improved by adopting the emasculation method of the gibberellic acid spraying of the 45-degree angle chaff in the oblique direction, the labor intensity and the work complexity of an operator are greatly reduced, and the problems of blocking of pollination channels also exist in the conventional finger-snapping water-spraying finger-popping method of the warm soup.
Example 3
Effect of different gibberellic acid dosage on outcrossing maturing rate
The test method comprises the following steps: selecting a rice variety as newly selected japonica 16-287, selecting a sorghum variety as local short-stalk glutinous sorghum soil, adopting an obliquely upward 45-degree angle glume shearing and emasculation method, setting the using amount of 3% gibberellic acid emulsifiable solution to be 4 different amounts of 200-time solution, 400-time solution, 600-time solution and 800-time solution for treatment, repeating the treatment for 3 times, performing emasculation for 10 ears each time, reserving 25 small ears each time, performing uniform spot spraying treatment by using gibberellic acid solutions with different concentrations, and checking the outcrossing maturing rate of each treatment 15 days after each treatment pollination.
TABLE 3 Effect of different amounts of gibberellic acid on outcrossing setting percentage
| Treatment of | Outcrossing maturing rate (%) |
| 200 times of 3% gibberellic acid emulsifiable solution | 0.27c |
| 400-fold liquid of 3% gibberellic acid emulsifiable concentrate | 0.87b |
| 600-fold liquid of 3% gibberellic acid emulsifiable solution | 1.20a |
| 3% gibberellic acid emulsifiable concentrate800 times liquid | 0.93ab |
It can be seen from table 3 that there is a great difference in the rate of heterozygosity of gibberellic acid with different dosages, the highest rate of heterozygosity of 2 times of the 3% gibberellic acid emulsifiable solution can be 1.20%, the rate of heterozygosity of 800 times of the 3% gibberellic acid emulsifiable solution and the rate of heterozygosity of 400 times of the 3% gibberellic acid emulsifiable solution are 0.93% and 0.87%, and the rate of heterozygosity of 200 times of the 3% gibberellic acid emulsifiable solution is only 0.27%, the analytic reason is that the low concentration gibberellic acid solution has the function of improving the vitality of pistil stigmas, while the high concentration gibberellic acid solution can accelerate the aging of pistil stigmas and has obvious damage effect on pistils, so that the 3% gibberellic acid emulsifiable solution is adopted for the castration of rice hybridization, and the 600 times of the solution should be diluted.
Example 4
The gibberellic acid spray castration method adopts different glume cutting modes to influence castration efficiency and outcrossing seed setting rate
The test method comprises the following steps: selecting a rice variety as newly selected japonica 16-287, selecting a sorghum variety as a local short-stalk glutinous sorghum soil variety, dividing a glume shearing mode into a conventional flat-mouth glume shearing method and a 45-degree oblique-upward glume shearing method, uniformly and pointly spraying 600 times of gibberellic acid solution to each spikelet after glume shearing is finished, repeating the treatment for 3 times, removing 10 spikes each time, keeping 25 spikes each time, accurately timing the castration time of each treatment in the next day, accurately timing the castration time to the second, comparing the treatment efficiency, removing impurities in each treatment, checking the outcrossing maturing rate 15 days after pollination, and comparing the influence of different glume shearing methods on the outcrossing maturing rate.
TABLE 4 influence of different glume-cutting modes of gibberellic acid spraying on castration efficiency and outcrossing seed setting rate
| Glume cutting mode | Emasculation efficiency (ear/second) | Outcrossing maturing rate (%) |
| Conventional glume cutting method with flat opening | 174.3a | 0.67b |
| Glume cutting method at 45-degree angle in oblique direction | 90.7b | 1.33a |
It can be seen from table 4 that the difference of the influence of different glume shearing methods on castration efficiency is significant, the castration efficiency of the glume shearing method by the 45-degree-angle oblique-upward glume shearing method is improved by 92.2% compared with that of the conventional horizontal-opening glume shearing method, and the analysis reason is that the horizontal-opening glume shearing method has a significant opening shearing area smaller than that of the 45-degree-angle oblique-upward glume shearing method, and after the gibberellic acid solution is sprayed, part of residual stamen pollen of the stamens expands after absorbing water to cause channel blockage, so that the residual stamens can not smoothly eject the small-ear glumes of the rice, and therefore, a long time is wasted for manual tasking treatment, the castration efficiency is affected, the damage probability of the stamens is increased, the outcrossing maturing rate is reduced, and the outcrossing maturing rate of the conventional horizontal-opening glume shearing method is only half of the 45-degree-angle oblique-upward glume shearing method.
Example 5
Influence of different collection times of sorghum pollen on rice sorghum outcrossing maturing rate
The test method comprises the following steps: selecting a rice variety as newly selected japonica 16-287, selecting a sorghum variety as local short-stalk glutinous sorghum soil, adopting an obliquely upward 45-degree glume cutting method, uniformly spraying 600 times of 3% gibberellic acid emulsifiable solution at a point after glume cutting of each ear is finished, and respectively collecting sorghum pollen for the following time: collecting sorghum blossoming ears at 6 am, putting the sorghum blossoming ears into a foam box with a wet tissue spread at the bottom, pollinating at about 30 minutes at 8 am, additionally, collecting the sorghum blossoming ears at about 8 am, pollinating at about 30 minutes at 8 am, repeating for 3 times every treatment, removing 10 ears every time, keeping 25 spikelets every ear, checking the outcrossing seed setting rate of each treatment 15 days after each treatment finishes pollination, and comparing the influence of different collection time of sorghum pollen on the outcrossing seed setting rate.
TABLE 5 Effect of different Collection time of sorghum pollen on outcrossing seed set
| Sorghum pollen collection time | Outcrossing maturing rate (%) |
| 6 am in the morning | 1.46a |
| 8 am | 0.53b |
As can be seen from Table 5, the difference of the different sorghum pollen collection times on the rice sorghum outcrossing maturing rate is obvious, the sorghum pollen outcrossing maturing rate collected at 6 am is 1.46%, the analysis reason is that the sorghum pollen collected at 8 am has a large amount of loose powder of the flowering sorghum at about 8 am, the sorghum pollen collection can cause loss of a large amount of sorghum pollen, the sorghum pollen intensity is insufficient during pollination, the outcrossing and fructification rate is low, sorghum pollen is collected about 6 a.m., although the sorghum blooms but a large amount of pollen is not scattered, the loss of the pollen is small when the sorghum pollen is collected, the collected sorghum ears are placed in a foam box with wet paper towels spread at the bottom, high-humidity environmental conditions required by sorghum pollen scattering can be simulated, the sorghum pollen can be scattered, and high pollen quantity is ensured when 30 minutes of pollination is carried out at 8 am, so that the outcrossing maturing rate is improved.
With reference to example 1, example 2, example 3, example 4 and example 5, the distant hybridization method for rice sorghum specifically comprises the following steps:
1) selecting target rice ears according to test requirements, reserving 20-25 small ears which can bloom the next day, and removing other small ears which already bloom and cannot bloom the next day.
2) And (4) carrying out glume cutting on the reserved spikelets at 1/2 along the middle part of the glume shell of each spikelet by adopting a 45-degree-angle upwards inclined glume cutting method, and paying attention to cutting anthers of each stamen without damaging pistils.
3) After each ear of rice is cut, uniformly spraying the small ears of rice with 600 times of 3% gibberellic acid emulsifiable solution, soaking each small ear in the chemical solution, and labeling.
4) The residual stamen anthers protruding the glumes were pinched off or flicked with fingers and covered with kraft paper bags 7 o' clock the next morning.
5) After the sorghum blooms every day, at about 6 am, cutting off the bloomed sorghum ears, putting the bloomed sorghum ears into a foam box with a wet tissue laid at the bottom, covering the foam box, slightly transporting the foam box to a paddy field, and pollinating the sorghum ears at about 30 minutes at 8 am.
6) The pollination method comprises the steps of gently taking down a large number of sorghum stalks which loose powder, inserting the sorghum stalks into a kraft paper bag, quickly rotating to achieve the purpose of uniformly scattering the powder, repeatedly pollinating for many times to ensure the pollination strength, and marking the hang-tag after pollination is finished.
The above description is intended to describe the preferred embodiments of the present invention in detail, but the embodiments are not intended to limit the scope of the claims, and all equivalent changes and modifications made within the technical spirit of the present invention should be covered by the present invention.
Claims (6)
1. A distant hybridization method for rice and sorghum is characterized by mainly comprising the steps of cutting glumes of small ears reserved for target rice ears in an obliquely upward glume cutting method in the previous day, marking the small ears, spraying a gibberellin solution with low concentration to each small ear at a uniform point after glume cutting of each ear is finished, manually pinching off or slightly flicking off residual stamen anthers extending out of glumes before the next 7 days, and timely collecting sorghum pollen and timely pollinating by adopting a certain technical means every day in order to improve the outcrossing maturing rate of rice and sorghum.
2. The method of claim 1, wherein glume is cut at 1/2 points along the middle of each tassel, the glume cutting angle is controlled at about 45 degrees, and the anther of each stamen is cut without damaging the pistil, so that the method can maximize the area of a glume pollination channel, increase the probability that sorghum pollen falls into the pistil, ensure that gibberellic acid solution can fully infiltrate into filaments of the rice stamen, ensure the drug effect of gibberellic acid solution, ensure that the anther of the stamen expands after the gibberellic acid solution is sprayed so as not to block the pollination channel, ensure that residual anther of the stamen can smoothly eject spikelet glumes, and cut the anther of the stamen so as to ensure that the pollen loses vigor after absorbing water, thereby improving the success rate of outcrossing.
3. The spraying of the low-concentration gibberellic acid solution as claimed in claim 1, which means that 3% gibberellic acid emulsifiable solution is diluted by 600 times, and the concentration can not only ensure that residual stamen anther can basically eject small ear glumes, but also improve the activity of rice pistil stigma and obviously improve the rice sorghum distant hybridization outcrossing maturing rate.
4. The method for timely collecting and pollinating sorghum pollen according to claim 1, wherein the flowering sorghum ears are collected at about 6 am every day, the loss of collected sorghum pollen is low, and the rice is pollinated repeatedly when about 30 minutes of sorghum pollen is sufficient for pollen scattering every 8 hours.
5. The method for collecting sorghum pollen according to claim 1, which is to put the cut sorghum ears into a foam box with wet paper towels laid at the bottom and cover the foam box, and aims to simulate the high-humidity environment required by sorghum pollen scattering and ensure that the sorghum pollen is sufficient during pollination.
6. The invention is not only applicable to the technical field of distant hybridization of rice and sorghum, but also belongs to the protection scope of the patent if the glume cutting and emasculation method adopted by the invention is applied to conventional rice and wheat hybridization breeding.
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