Scientia Agricultura Sinica ›› 2023, Vol. 56 ›› Issue (7): 1275-1282.doi: 10.3864/j.issn.0578-1752.2023.07.006

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• SPECIAL FOCUS: PANICLE DEVELOPMENT AND YIELD BREEDING IN RICE • Previous Articles     Next Articles

Expression Pattern of the Rice α-Amylase Genes Related with the Process of Floret Opening

ZHANG Ji(), ZHOU ShangLing, HE Fa, LIU LiSha, ZHANG YuJuan, HE JinYu, DU XiaoQiu()   

  1. Nanchong Academy of Agricultural Sciences, Nanchong 637000, Sichuan
  • Received:2022-10-31 Accepted:2023-01-19 Online:2023-04-01 Published:2023-04-03

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Abstract:

【Objective】 Starch degradation is involved in lodicule absorbing abundant water and swelling during rice floret opening, but the amylase genes associated with this process have not been identified yet. 【Method】 To identify the swelling of tissues during floret opening, the in vitro rice panicles absorbed diluted Fuchsin basic and the dye remains were observed after florets were closing again. The starch grain distribution in rice florets before and during anthesis from stage 11 to stage 14 (according to 14 stages of rice anther development) was detected using iodide staining. The spatial-temporal expression patterns of 10 α-amylase genes were detected by RT-PCR, RT-qPCR and GUS staining. 【Result】 Before floret opening, the stamens, pistils and lodicules are enclosed by the lemma and palea through marginal tissues of palea (mtp). Rapid swelling of the lodicules causes floret opening by separating the lemma from the palea. After the in vitro panicles absorbed diluted Fuchsin basic during floret opening, the dye remains were observed located in the joint between mtp and lodicules and filaments. Iodide staining showed that the starch grains were mainly located in the stamens and mtp and a small amount of starch grains in the lodicules at stage 12 (before floret opening), whereas the starch grains in the mtp and lodicules were almost completely degraded at stage 13-14 (during floret opening). RT-PCR showed that OsRAmy2A and OsRAmy3D began to express from stage 12 and were expressed with high levels at stage 13-14. The expression levels of the two genes decreased at DAP1 (1 day after pollination). OsRAmy3E and OsRAmy3F kept expressed during this process. The expression level of OsRAmy3E was higher than that of OsRAmy3F. The RT-qPCR analysis showed that the expression level of OsRAmy2A increased most dramatically at stage 13-14, followed by OsRAMy3A and OsRAMy3E. Further, the transgenic plants expressing the GUS reporter gene driven by the OsRAmy2A promoter were generated. The GUS signaling was located only in the lemma, palea and mtp at stage 12 and the expression of the GUS gene driven by the RAmy2A promoter was induced in the mtp, lodicules and filaments at stage 13-14. 【Conclusion】 These data indicated that starch grain degradation in the mtp and lodicules at stage 13-14 might be related with high expression levels of some α-amylase genes such as OsRAmy2A and OsRAmy3D, probably involved in controlling lodicules swelling and floret opening in rice.

Key words: rice, α-amylase, floret opening, starch grain, lodicules swelling

Fig. 1

The phenotype of rice floret opening A: Stage 13 (Left), stage14 (Right); Bar=2 mm; B: Imagination of lodicules at stage 13, Bar=0.5 mm; C: Imagination of lodicules at stage 14, Bar=0.5 mm"

Fig. 2

Absorption and remains of diluted Fuchsin basic during floret opening A: The joint between mtp and lodicules; B: Filaments, Bar=2 mm. Lo: Lodicules; mtp: Marginal tissues of palea; P: Palea; Fi: Filaments. The same as below"

Fig. 3

Lodine staining reaction of rice florets during 11-14 stages A-B: Stage 11, A, magnification of B with lemma removed; C-D: Stage 12, C, magnification of D; E: Stage 13-14. Bar=2 mm. L: Lemma; S: Stamen; R: Rachilla. The same as below "

Fig. 4

The analysis of 10 α-amylase genes before and during rice floret opening by RT-PCR DAP1: 1 day after pollination. The numbers in parentheses are cycles of RT-PCR for the genes"

Fig. 5

The analysis of 4 α-amylase genes before and during rice floret opening by RT-qPCR"

Fig. 6

The expression profiling of the OsRAmy2A Promotor::GUS transgenic rice in florets before and during anthesis by GUS staining A-C: Stage 11; D-F: Stage 12; G-I: Stage 13-14, Bar=2 mm. An: Anther"

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