赤霉素对‘苏翠1号’梨花芽松散的影响

doi:10.16420/j.issn.0513-353x.2024-0810

摘要/Abstract

摘要：

以‘苏翠1号’梨花芽为试材，比较正常花芽和松散花芽的赤霉素含量，转录组比较分析赤霉素合成途径相关基因表达情况，研究施加赤霉素抑制剂多效唑控制花芽松散的效果，探讨花芽松散与赤霉素之间的关系。结果表明，松散花芽中赤霉素含量显著高于正常花芽，其中GA₁、GA₃、GA₄和GA₇含量是正常花芽的1.25 ~ 2.87倍。转录组分析显示松散花芽与正常花芽有3 290个差异表达基因，松散花芽中上调表达的2 046个，下调表达的1 244个。qRT-PCR结果表明，松散花芽中GA2ox1、GA2ox2、GA3ox、GA20ox和GID1基因上调表达，GAI、GASA3/4/6/14、GID2和KAO下调表达，并且与转录组测序结果一致。施加多效唑后一年生枝条腋花芽中赤霉素含量减少，花芽松散比例下降，赤霉素含量与花芽松散比例呈显著正相关（r = 0.94）。喷施多效唑可抑制‘苏翠1号’梨腋花芽中GA2ox1/2、GA3ox、GA20ox、GASA3/4/6/14、GID1和KAO的转录水平，促进GAI和GID2表达，从而调控赤霉素的合成代谢。研究结果表明生长季赤霉素的异常升高会导致梨花芽松散，喷施多效唑通过调控合成途径相关基因表达，抑制赤霉素合成，减轻梨花芽的松散程度。

关键词: 梨, 花芽松散, 转录组, 赤霉素, 基因表达

Abstract:

To explore the relationship between flower buds wizening and gibberellin，the flower bud from Pyrus pyrifolia cultivar‘Sucui 1’was selected as the experimental material in this study. The gibberellin contents of‘Sucui 1’pear were compared between normal flower buds and wizened flower buds，and the expression of genes related to the gibberellin synthesis pathway once flower buds wizening were analyzed. Then the effect of applying gibberellin inhibitor paclobutrazol to control flower buds wizening was studied in the growing season. The results showed that there was significant difference in gibberellin contents between normal flower buds and wizened flower buds. The contents of GA₁，GA₃，GA₄ and GA₇ in wizened flower buds were 1.25-2.87 times of those in normal flower buds. Transcriptome analysis showed that，comparing with normal flower buds，wizened flower buds had 3 290 differentially expressed genes in which 2 046 were up-regulated and 1 244 were down-regulated. qRT-PCR confirmed the up-regulated expression of GA2ox1（gibberellin 2-oxidas 1），GA2ox2 （gibberellin 2-oxidas 2），GA3ox（gibberellin 3-oxidas），GA20ox （gibberellin 20-oxidas）and GID1（gibberellin insensitive Dwarf1）genes in wizened flower buds. Meanwhile，GAI（gibberrellic acid insensitive），GASA3/4/6/14（Arabidopsis gibberrellic acid stimulating protein 3，4，6 and 14），GID2（gibberellin Insensitive Dwarf2）and KAO（ent-kaurenoic acid monooxygenase）were down-regulated in wizened flower buds. And their expression information was consistent with the results of transcriptome sequencing. After the application of polybulobuzole，the gibberellin contents in axillary flower buds of annual branches reduced and the proportion of wizened flower buds in‘Sucui 1’decreased in mid-September. The content of gibberellin was significantly positively correlated with the proportion of wizened flower buds（r = 0.94）. Furthermore，the transcription levels of GA2ox1/2，GA3ox，GA20ox，GAI，GASA3/4/6/14，GID1/2 and KAO in‘Sucui 1’flower bud were affected by applying polybulobuzole，which took part in the synthesis and metabolism of gibberellin. Spraying paclobutrazol can inhibit the transcription levels of GA2ox1/2，GA3ox，GA20ox，GASA3/4/6/14，GID1，and KAO in the axillary flower buds of‘Sucui 1’pear，but promote the expression of GAI and GID2，furthermore regulate the synthesis and metabolism of gibberellin. The research results indicate that an abnormal increase in gibberellin during the growing season can lead to wizened flower buds in pear. Spraying paclobutrazol would regulate the expression of genes related to the gibberellin synthesis pathway，inhibit it synthesis， and relief the wizened degree of pear flower buds.

Key words: pear, flower bud wizening, transcriptome, gibberellin, gene expression

李慧, 阚家亮, 李晓刚, 王中华, 蔺经, 杨青松. 赤霉素对‘苏翠1号’梨花芽松散的影响[J]. 园艺学报, 2025, 52(10): 2713-2726.

LI Hui, KAN Jialiang, LI Xiaogang, WANG Zhonghua, LIN Jing, YANG Qingsong. Effect of Gibberellin on Wizened Flower Buds Formation of Pyrus pyrifolia Cultivar‘Sucui 1’During the Growing Season[J]. Acta Horticulturae Sinica, 2025, 52(10): 2713-2726.

图/表 9

图1 ‘苏翠1号’正常花芽和松散花芽形态比较

Fig. 1 Morphology comparison between normal flower buds and wizened flower buds of Pyrus pyrifolia‘Sucui 1’

表1 实时荧光定量PCR引物

Table 1 Primers used for qRT-PCR in this study

基因名称 Gene name	正向引物序列（5′-3′） Forward primer sequence	反向引物序列（5′-3′） Reverse primer sequence	产物长度/bp Product length
Actin7（GWHGBAOS001891）	AATGAACTTCGTGTTGCTCCTG	CACCTGAGTCCAGCACAATACC	171
GA2ox1（GWHGBAOS003456）	GACCTCACAAAACCAGACTCCA	TCAACCCAACCAACATCACC	230
GA2ox2（GWHGBAOS018799）	AAGGTCATCAACCACGGCG	CCTGCTCTTTGCTTCTCGGA	101
GA20ox（GWHGBAOS016723）	GTGCCTCAGTTCTCTTGTCC	GAGCAGCAACCAAGTGTCGT	168
GA3ox（GWHGBAOS016475）	TGGCAGATCC CTATAGAGCT	TCAAATTCCACATCCTCCAT	266
GAI （GWHGBAOS021707）	GGTTCACGGAGGCATTGCATT	TCGGTCCCCACCTTCACAT	143
GASA3（GWHGBAOS018317）	CTTGTTGTGGTGGTTTTGTT	AGCCTTCTGTTTGCTCCTTTC	107
GASA4（GWHGBAOS038046）	CAGCCAGACGCAGTACCACA	AACCCAGGGGGTACACACAG	81
GASA6（GWHGBAOS011991）	CGGAATCAGTTAGACAGTGG	TTGTGATGGGCATTGGTAGC	66
GASA14（GWHGBAOS037916）	CCGCTAGGGCTTCATCACTT	CGCTGGAGCCTTGGATAGAG	96
GID1（GWHGBAOS006870）	TTACCAGTTTGCGCCCCA	CCGTGGAAAAAGATTATGACCG	102
GID2（GWHGBAOS036864）	GCGGCGTCCTCCTCTGATTC	GAAAAGCGAGAGCGACAGATTC	116
KAO（GWHGBAOS022579）	CGCAGAAGGGCTTGACACT	AGAGTTTCATCAATCACCTGGC	110

表2 ‘苏翠1号’花芽赤霉素含量

Table 2 Content of gibberellin in flower buds of Pyrus pyrifolia‘Sucui 1’

花芽类型 Flower bud type	赤霉素含量/（ng · g^-1）Gibberellin content
花芽类型 Flower bud type	GA₁	GA₃	GA₄	GA₇
正常花芽 Normal flower bud	2.81 ± 0.47	1.35 ± 0.04	0.22 ± 0.02	0.06 ± 0.01
松散花芽 Wizened flower bud	8.07 ± 0.40 ^**	1.69 ± 0.10 ^**	0.42 ± 0.04 ^**	0.11 ± 0.01 ^**

图2 ‘苏翠1号’正常花芽和松散花芽前10个KEGG通路中差异表达基因热图

Fig. 2 Heat map of differentially expressed genes in the first 10 KEGG pathway between normal and wizened flower buds of Pyrus pyrifolia ‘Sucui 1’

图3 ‘苏翠1号’正常花芽和松散花芽差异表达基因参与赤霉素合成和信号转导KEGG图绿色表示下调，红色表示上调；DELLA：DELLA蛋白；GA2ox：赤霉素2-氧化酶；GA20ox：赤霉素20-氧化酶；GA3ox：赤霉素3-氧化酶；GID1：赤霉素不敏感型Dwarf1；GID2：赤霉素不敏感型Dwarf2；KAO：内—贝壳杉烯酸单加氧酶；P450：细胞色素P450单加氧酶

Fig. 3 KEGG map of differentially expressed genes involved in gibberellin synthesis and signal transduction in normal and wizened flower buds of Pyrus pyrifolia‘Sucui 1’ Green indicates down-regulated expression，and red indicates up-regulated expression；DELLA：DELLA protein；GA2ox：Gibberellin 2-oxidase；GA20ox：Gibberellin 20-oxidase；GA3ox：Gibberellin 3-oxidase；GID1：Gibberellin insensitive Dwarf1；GID2：Gibberellin insensitive Dwarf2；KAO：Ent-kaurenoic acid monooxygenase；P450：Cytochrome P450 monooxygenase

图4 ‘苏翠1号’不同花芽中赤霉素相关基因FPKM值和相对表达水平 **代表在P < 0.01水平有显著性差异（Student’s t-test）；柱型图为基因相对表达量，红线表示转录组测序获得的FPKM值；GA2ox：赤霉素2-氧化酶；GA20ox：赤霉素20-氧化酶；GA3ox：赤霉素3-氧化酶；GAI：赤霉酸不敏感；GASA：拟南芥赤霉酸刺激蛋白；GID：赤霉素不敏感型Dwarf；KAO：内—贝壳杉烯酸单加氧酶

Fig. 4 FPKM values and relative expression levels of gibberellin-related genes in different types of flower buds from Pyrus pyrifolia‘Sucui 1’ ** indicates significant difference at P < 0.01 level（Student’s t-test）；The histogram shows the relative expression level of genes，and the red line shows the FPKM value obtained by transcriptome sequencing；GA2ox：Gibberellin 2-oxidase；GA20ox：Gibberellin 20-oxidase；GA3ox：Gibberellin 3-oxidase；GAI：Gibberrellic acid insensitive；GASA：Gibberellic acid-timulated Arabidopsis；GID：Gibberellin insensitive Dwarf；KAO：ent-Kaurenoic acid monooxygenase

表3 多效唑喷施处理‘苏翠1号’花芽情况

Table 3 The information of flower buds from Pyrus pyrifolia‘Sucui 1’after spraying paclobutrazole

多效唑/ （mg · L^-1） Paclobutrazol	调查花芽总数 The total number of investigating flower buds	赤霉素含量/（ng · g^-1） Gibberellin content （GA₁+ GA₃+ GA₄ + GA₇）	正常花芽数 Number of normal flower buds	松散花芽Wizened flower buds
多效唑/ （mg · L^-1） Paclobutrazol	调查花芽总数 The total number of investigating flower buds	赤霉素含量/（ng · g^-1） Gibberellin content （GA₁+ GA₃+ GA₄ + GA₇）	正常花芽数 Number of normal flower buds	数量 Number	比例/% The proportion
0（清水对照 Water control）	500	6.96 ± 0.44 A	249	251	50.2 A
10	500	5.92 ± 0.40 B	259	241	48.2 B
20	500	5.02 ± 0.34 C	302	198	39.6 C
30	500	3.79 ± 0.26 D	399	101	20.2 D

图5 喷施不同浓度多效唑后‘苏翠1号’一年生枝条腋花芽情况

Fig. 5 Axillary flower buds of annual branches from Pyrus pyrifolia‘Sucui 1’after spraying with different concentrations polyphobulozole

图6 喷施不同浓度多效唑后‘苏翠1号’花芽中赤霉素相关基因的相对表达水平不同小写字母代表在P < 0.05水平有显著性差异（邓肯多范围测试）；GA2ox：赤霉素2-氧化酶；GA20ox：赤霉素20-氧化酶；GA3ox：赤霉素3-氧化酶；GAI：赤霉酸不敏感；GASA：拟南芥赤霉酸刺激蛋白；GID：赤霉素不敏感型Dwarf；KAO：内-贝壳杉烯酸单加氧酶；基因ID：GID1（GWHGBAOS006870）、GID2（GWHGBAOS036864）、GA2ox1（GWHGBAOS003456）、GA2ox2（GWHGBAOS018799）、GA3ox（GWHGBAOS016475）、GA20ox（GWHGBAOS016723）、GAI（GWHGBAOS021707）、GASA3（GWHGBAOS018317）、GASA4（GWHGBAOS038046）、GASA6（GWHGBAOS011991）、GASA14（GWHGBAOS037916）、KAO（GWHGBAOS022579）

Fig. 6 Effect of spraying polybulobuzole on relative expression levels of gibberellin-related genes in flower buds from Pyrus pyrifolia‘Sucui 1’ Different lowercase letters represent significant differences at the P < 0.05 level（Duncan's multiple range test）；GA2ox：Gibberellin 2-oxidase；GA20ox：Gibberellin 20-oxidase；GA3ox：Gibberellin 3-oxidase；GAI：Gibberrellic acid insensitive；GASA：Gibberellic acid-timulated Arabidopsis；GID：Gibberellin insensitive Dwarf；KAO：Ent-kaurenoic acid monooxygenase；Gene ID：GID1（GWHGBAOS006870），GID2（GWHGBAOS036864）、GA2ox1（GWHGBAOS003456），GA2ox2（GWHGBAOS018799），GA3ox（GWHGBAOS016475），GA20ox（GWHGBAOS016723），GAI（GWHGBAOS021707），GASA3（GWHGBAOS018317），GASA4（GWHGBAOS038046），GASA6（GWHGBAOS011991），GASA14（GWHGBAOS037916），KAO（GWHGBAOS022579）

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