Method for regulating heading stage of rice and application thereof

Abstract

The invention relates to a method for regulating and controlling the heading stage of rice and application of the method, and belongs to the technical field of plant biology. The invention provides a method for regulating and controlling the heading period of rice. Red light is adopted to irradiate rice plants for 3-10 min. The method provided by the invention proves that the heading period can be regulated and controlled by carrying out short-time red light irradiation treatment on the rice plants under a dark treatment condition, and the method has wide applicability to different varieties of rice plants, so that a theoretical foundation is laid for regulating and controlling the heading period of the rice through the red light irradiation treatment; and the method has important practical significance for researching the heading period of the rice regulated by light quality and solving the problem of different hybrid flowering periods of different varieties of rice.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of plant biotechnology, and particularly relates to a method for regulating the heading stage of rice and application thereof. BACKGROUND

[0002] Rice is an important food crop, and the guarantee of its yield is of great significance to national food security. Heterosis of rice is widely used in actual production, but in the process of hybrid breeding, the mismatch of flowering period is an important factor leading to the failure of normal hybridization of rice, resulting in low seed setting rate or even failure of seed production. The reasons for the mismatch of flowering period mainly include the difference in the perception of environmental factors between rice parents, irregular climate change, extreme weather factors, and different artificial cultivation and management measures.

[0003] At present, the methods for regulating the heading stage of rice mainly include density regulation, seedling age regulation, fertilizer regulation, water regulation, multi-effect azole regulation, and LED light quality regulation, but the above methods generally rely on manual experience and have the defects of high cost and difficult to control, so the existing technology urgently needs a simple and efficient technical solution for regulating the heading stage of rice. SUMMARY

[0004] The purpose of the present application is to provide a method for regulating the heading stage of rice and application thereof, so as to solve the problems of high energy consumption, large subjective influencing factors, and difficult to accurately control in the prior art.

[0005] In order to achieve the above-mentioned purpose of the application, the present application provides the following technical solutions: The present application provides a method for regulating the heading stage of rice, which adopts red light to irradiate the rice plant for 3-10 minutes.

[0006] Preferably, the wavelength of the red light is 625-680 nm.

[0007] Preferably, the culture temperature of the rice is 25-30 DEG C, the culture time of the rice is 21-30 days, the light treatment time of the rice is 8-12 hours per day, and the dark treatment time of the rice is 14-16 hours per day.

[0008] Preferably, the rice is in the heading stage.

[0009] Preferably, the irradiation treatment time period is 1-14 hours of dark treatment.

[0010] Preferably, the duration of the irradiation treatment is more than one week.

[0011] Preferably, the temperature for culturing after irradiation treatment is 25–30°C, the light treatment time for culturing after irradiation treatment is 8–12 h / d, and the dark treatment time for culturing after irradiation treatment is 14–16 h / d.

[0012] The present invention also provides the application of the method in delaying the heading period of rice.

[0013] The present invention has the following technical effects and advantages: This invention demonstrates that short-term red light irradiation of rice plants under dark conditions can regulate the heading stage, especially that red light irradiation for 3–10 minutes at the 7th hour of dark treatment can inhibit flowering hormones. Hd3a The expression level of genes can be adjusted, which can delay the heading period of rice plants. Moreover, 3 minutes of red light irradiation can achieve the same technical effect as 10 minutes of irradiation. This shows that short-term red light irradiation of rice plants under dark conditions can play an important role in regulating the heading period of rice, which is of great significance for the study of light quality regulation of the heading period. This invention also demonstrates that red light irradiation of rice plants under dark conditions delays the heading period by different durations; the longer the duration, the longer the delay. This indicates that the duration of red light irradiation under dark conditions is closely related to the heading period of rice, and that it has broad applicability to different rice varieties. This lays a theoretical foundation for regulating the heading period of rice with red light irradiation and has significant practical implications for research on light quality regulation of the heading period of rice and for solving the problem of asynchronous flowering in hybrid rice varieties. Attached Figure Description

[0014] Figure 1 To investigate the effects of 10 min red light irradiation on rice seedlings of the Tokyo variety at different time points during the dark treatment. Hd3a Gene transcription level and heading results; where A represents the results of 10 min red light irradiation on rice seedlings of the Tokyo variety at different time points during the dark treatment. Hd3a Gene transcription level: B is the heading phenotype of rice seedlings of Tokyo variety at the heading stage after 3 weeks of 10 min red light irradiation at ZT11; C is the statistical graph of heading results of rice seedlings of Tokyo variety at the heading stage after 3 weeks of 10 min red light irradiation at ZT11; D is the heading phenotype of rice seedlings of Tokyo variety at the heading stage after 3 weeks of 10 min red light irradiation at ZT17; E is the statistical graph of heading results of rice seedlings of Tokyo variety at the heading stage after 3 weeks of 10 min red light irradiation at ZT17. Figure 2Fig. 1 shows the heading results of rice seedling materials of Tokyo variety at the heading stage after being treated with 10 min red light irradiation for different durations at ZT17; wherein A is a heading phenotype diagram of rice seedling materials of Tokyo variety at the heading stage after being treated with 10 min red light irradiation for different durations at ZT17, and B is a statistical diagram of the heading results of rice seedling materials of Tokyo variety at the heading stage after being treated with 10 min red light irradiation for different durations at ZT17; Figure 3 Fig. 2 shows the heading results of rice seedling materials of Tokyo variety at the heading stage after being treated with 3 min red light irradiation for different durations at ZT17; wherein A is a heading phenotype diagram of rice seedling materials of Tokyo variety at the heading stage after being treated with 3 min red light irradiation for different durations at ZT17, and B is a statistical diagram of the heading results of rice seedling materials of Tokyo variety at the heading stage after being treated with 3 min red light irradiation for different durations at ZT17; Figure 4 Fig. 3 shows the heading results of rice seedling materials of Zhonghua 11 and Tokyo varieties at the heading stage after being treated with 10 min red light irradiation for 3 weeks at ZT17; wherein A is a heading phenotype diagram of rice seedling materials of Zhonghua 11 variety at the heading stage after being treated with 10 min red light irradiation for 3 weeks at ZT17, B is a heading result diagram of rice seedling materials of Zhonghua 11 variety at the heading stage after being treated with 10 min red light irradiation for 3 weeks at ZT17, C is a heading phenotype diagram of rice seedling materials of Tokyo variety at the heading stage after being treated with 10 min red light irradiation for 3 weeks at ZT17, and D is a heading result diagram of rice seedling materials of Tokyo variety at the heading stage after being treated with 10 min red light irradiation for 3 weeks at ZT17. DETAILED DESCRIPTION

[0015] The application provides a method for regulating the heading stage of rice, which comprises treating the rice plant with red light irradiation for 3-10 min, preferably 3 min.

[0016] In the application, the wavelength of the red light is 625-680 nm, preferably 660 nm.

[0017] In the application, the culture temperature of the rice is 25-30°C, preferably 28°C; the culture time of the rice is 21-30 d, preferably 28 d; the light treatment time of the rice is 8-12 h / d, preferably 10 h / d; and the dark treatment time of the rice is 14-16 h / d, preferably 14 h / d.

[0018] In the application, the rice is at the heading stage.

[0019] In the application, the irradiation treatment time is 1-14 h, preferably 7 h, of the dark treatment.

[0020] In the present application, the duration of the irradiation treatment is more than 1 week, preferably 3 weeks.

[0021] In the present application, the temperature of the post-irradiation culture is 25-30°C, preferably 28°C; the light treatment time of the post-irradiation culture is 8-12 h / d, preferably 10 h / d; and the dark treatment time of the post-irradiation culture is 14-16 h / d, preferably 14 h / d.

[0022] The present application also provides the use of the method in delaying the heading stage of rice.

[0023] The technical solutions provided by the present application will be described in detail below in conjunction with the examples, but they should not be understood as limiting the scope of protection of the present application.

[0024] The test rice varieties of the present application are Tokyo (DJ), Zhonghua 11 (ZH11) and Nipponbare (Nip), and the seeds of each rice variety are prepared by the present laboratory; In the reagent of the present application, SYBR ® Green Realtime PCR Master Mix reagent is purchased from Tokyo Biochemical (Shanghai) Co., Ltd.; In the instrument of the present application, Applied Biosystems TM QuantStudio 3 real-time fluorescent quantitative PCR instrument is purchased from American ABI Company.

[0025] Example 1: Cultivation of rice seedling materials Select well-developed and plump Tokyo (DJ), Zhonghua 11 (ZH11) and Nipponbare (Nip) rice seeds, soak them in distilled water at 37°C for 48 h, replace the water every 12 h during the soaking, and then continue to germinate in a humid environment for 24 h to obtain germinated seeds; cultivate the germinated seeds under short-day conditions of 28°C, 10 h light / 14 h darkness to the 21st day, select the rice seedlings with consistent growth, transplant them into 32-hole rice culture pots, and continue to cultivate for 7 days to obtain the heading stage rice seedling materials of each variety.

[0026] Example 2: Treatment of Tokyo rice seedling materials Rice seedlings of the Tokyo variety obtained in Example 1 at the heading stage were treated with 660 nm red light for 10 min at different time points of dark treatment (ZT11, ZT13, ZT15, ZT17, ZT19, ZT21, ZT23, corresponding to the 1 h, 3 h, 5 h, 7 h, 9 h, 11 h, and 13 h of dark treatment, respectively). They were then cultured under short-day conditions of 28℃, 10 h light / 14 h dark until the next day, and rice leaves were collected from each treatment. Rice seedlings of the Tokyo variety at the heading stage without red light treatment were used as a control. RNA was extracted from the rice leaves and reverse transcribed into cDNA. The floraltin gene was then analyzed. Hd3a qPCR primers Hd3a qPCR was performed using -qF / R, and rice was used as a substrate. Ubiquitin The nucleotide sequences of each primer, used as an internal reference gene, are shown in Table 1. The qPCR reaction program was as follows: 95℃ 2min → (95℃ 15s → 55℃ 15s → 72℃ 15s) × 40 cycles → 95℃ 15s → 60℃ 1min → 95℃ 15s. The exported qPCR data were analyzed. Hd3a gene CT value minus Ubiquitin The difference obtained from gene CT values ​​is used for 2-Δ CT calculations involve repeatedly averaging the results from the three techniques. Hd3a Gene expression levels; results as follows Figures 1 to 3 As shown.

[0027] Table 1 Nucleotide sequences of each qPCR primer The results showed that red light irradiation treatment of Tokyo variety rice seedlings at different times of night had an effect on... Hd3a Gene transcription levels showed varying degrees of repression, with red light irradiation treatment of ZT17 showing the greatest impact. Hd3a The transcriptional level of the gene is most strongly suppressed.

[0028] Example 2: Treatment of seedling materials of Zhonghua 11 and Nipponbare rice varieties Rice seedlings of the Zhonghua 11 and Nipponbare varieties obtained in Example 1 were treated with 660 nm red light for 10 min at the 7th hour of dark treatment (ZT17), and this treatment was continued for 3 weeks. Untreated Zhonghua 11 and Nipponbare rice seedlings at the heading stage were used as a control. The seedlings were then cultured under short-day conditions of 28℃, 10 h light / 14 h dark until heading was achieved. The heading time of each seedling was recorded. The results are as follows: Figure 4 As shown.

[0029] The results show that the red light irradiation treatment at night can effectively delay the heading stage of the rice seedling materials of Zhonghua 11 and Nipponbare varieties.

[0030] From the above examples, the present application provides a method for regulating the heading stage of rice and its application. The present application proves that short-time red light irradiation treatment on rice plants under dark treatment conditions can regulate the heading stage, and has wide applicability for different varieties of rice plants, thereby laying a theoretical foundation for red light irradiation treatment to regulate the heading stage of rice, and having important practical significance for the research of light quality regulating the heading stage of rice and solving the problem of different varieties of rice hybrid flowering period.

[0031] The above only describes the preferred embodiments of the present application, and it should be noted that for ordinary skilled persons in the art, several improvements and refinements can be made without departing from the principles of the present application, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A method for regulating the heading stage of rice, characterized in that, Rice plants were irradiated with red light for 3–10 minutes.

2. The method according to claim 1, characterized in that, The wavelength of the red light is 625–680 nm.

3. The method according to claim 2, characterized in that, The rice was cultivated at a temperature of 25–30℃ for 21–30 days, with a light treatment time of 8–12 h / d and a dark treatment time of 14–16 h / d.

4. The method according to claim 3, characterized in that, The rice is in the heading stage.

5. The method according to claim 4, characterized in that, The irradiation treatment period is from the first to the 14th hour of the dark treatment.

6. The method according to claim 5, characterized in that, The duration of the irradiation treatment is more than one week.

7. The method according to claim 6, characterized in that, The temperature for culturing after irradiation treatment is 25–30°C, the light treatment time for culturing after irradiation treatment is 8–12 h / d, and the dark treatment time for culturing after irradiation treatment is 14–16 h / d.

8. The application of the method according to any one of claims 1 to 7 in delaying the heading period of rice.

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