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Citation:

18 miRNAs in Larix kaempferi During Seed Germination

  • Received Date: 2013-08-24
  • The seed of Larix kaempferi (Siebold. & Zucc.) Gordon was used as material to investigate the potential molecular regulation mechanisms of miRNA during seed germination. The germination process of L.kaempferi seed includes the following: The imbibition of dry seed was completed after 24 hours of socking; the testa rupture occurred after 5 days and followed by radicle emergence (videlicet, endosperm rupture) after 9 days of germination culture; the cotyledons expanded fully after 28 days. The expression level of 18 miRNAs was relatively quantified using qRT-PCR technology among five key seed germinating stages, which including dry seed (0 day after germinating), imbibed seed (1st day after germinating), testa rupture (5th day), radicle emergence (9th day) and cotyledon extension (28th day). The results are as follows. (1)The expression levels of 18 miRNAs were the highest at dry seed stage and then went down rapidly at imbibed seed and testa rupture stage. The miRNAs were highly expressed in dry seed stage than in subsequent stages. The expression level in dry seed stage was 4.8 to 43.5 times, 17.2 to 1 000 times, 45.5 to 1 000 times and 62.5 to 1 862.2 times that of those in imbibed seed, testa rupture, radicle emergence and extension cotyledon stage, respectively. (2)The expression level of miR894 was the highest and that of miR408 was the lowest in each stage; and the others fluctuated, ranking from high to low as miR951, miR950, miR156, miR159, miR396, miR398, miR390, miR160, miR947, miR165, miR319, miR162, miR171, miR397, miR395 and miR172. (3)The target genes and their possible functions predicted using bioinformatics methods were responsible for the expression of a great number of binding proteins, cellular components and MAP kinase proteins associated with seed germination. (4) JR160237 was the target gene of miR160, and the cleaved fragments mapped exactly to the predicted cleavage site (between nucleotides 10 and 11 from the 5'end of miR160). JR160237 mRNA reached the peak at imbibed seed stage, then decreased and dynamically expressed at following stages with small fluctuation.
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18 miRNAs in Larix kaempferi During Seed Germination

  • 1. Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China
  • 2. Yunnan Academy of Forestry, Yunnan Key Laboratory for Forest Plant Cultivation and Utilization, Kunming 650201, Yunnan, China
  • 3. Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing 100091, China

Abstract: The seed of Larix kaempferi (Siebold. & Zucc.) Gordon was used as material to investigate the potential molecular regulation mechanisms of miRNA during seed germination. The germination process of L.kaempferi seed includes the following: The imbibition of dry seed was completed after 24 hours of socking; the testa rupture occurred after 5 days and followed by radicle emergence (videlicet, endosperm rupture) after 9 days of germination culture; the cotyledons expanded fully after 28 days. The expression level of 18 miRNAs was relatively quantified using qRT-PCR technology among five key seed germinating stages, which including dry seed (0 day after germinating), imbibed seed (1st day after germinating), testa rupture (5th day), radicle emergence (9th day) and cotyledon extension (28th day). The results are as follows. (1)The expression levels of 18 miRNAs were the highest at dry seed stage and then went down rapidly at imbibed seed and testa rupture stage. The miRNAs were highly expressed in dry seed stage than in subsequent stages. The expression level in dry seed stage was 4.8 to 43.5 times, 17.2 to 1 000 times, 45.5 to 1 000 times and 62.5 to 1 862.2 times that of those in imbibed seed, testa rupture, radicle emergence and extension cotyledon stage, respectively. (2)The expression level of miR894 was the highest and that of miR408 was the lowest in each stage; and the others fluctuated, ranking from high to low as miR951, miR950, miR156, miR159, miR396, miR398, miR390, miR160, miR947, miR165, miR319, miR162, miR171, miR397, miR395 and miR172. (3)The target genes and their possible functions predicted using bioinformatics methods were responsible for the expression of a great number of binding proteins, cellular components and MAP kinase proteins associated with seed germination. (4) JR160237 was the target gene of miR160, and the cleaved fragments mapped exactly to the predicted cleavage site (between nucleotides 10 and 11 from the 5'end of miR160). JR160237 mRNA reached the peak at imbibed seed stage, then decreased and dynamically expressed at following stages with small fluctuation.

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