Effect of Gibberellin on Wizened Flower Buds Formation of Pyrus pyrifolia Cultivar‘Sucui 1’During the Growing Season

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

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (10): 2713-2726.doi: 10.16420/j.issn.0513-353x.2024-0810

• Cultivation·Physiology & Biochemistry • Previous Articles Next Articles

Effect of Gibberellin on Wizened Flower Buds Formation of Pyrus pyrifolia Cultivar‘Sucui 1’During the Growing Season

LI Hui¹, KAN Jialiang¹^,², LI Xiaogang¹^,^*(), WANG Zhonghua¹, LIN Jing¹, YANG Qingsong¹

¹ Institute of Pomology，Jiangsu Academy of Agricultural Sciences/Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement，Nanjing 210014，China
² State Key Laboratory of Subtropical Silviculture，Zhejiang A & F University，Hangzhou 311300，China

Received:2024-11-26 Revised:2025-06-03 Online:2025-10-25 Published:2025-10-28
Contact: LI Xiaogang

Abstract

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

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.

Figures/Tables 9

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

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

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 ^**

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’

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

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

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

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

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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Effect of Gibberellin on Wizened Flower Buds Formation of Pyrus pyrifolia Cultivar‘Sucui 1’During the Growing Season

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