Expression Analysis of B-class MADS-box Genes from Catalpa speciosa

doi:10.11983/CBB15053

Abstract

Abstract: The cDNA sequence of B-class MADS-box genes CaspAP3 and CaspPI involved in flower development were isolated from the flower bud of Catalpa speciosa by homology cloning. The open reading frame (ORF) of CaspAP3 was 696 bp, encoding 231 amino acids, and the ORF of CaspPI was 639 bp, encoding 212 amino acids. Molecular phylogeny analysis and protein sequence alignment suggested that CaspAP3 groups with the AP3/DEF lineages and the C domain of CaspAP3 contained two conserved motifs, euAP3 and PI-derived motifs. However, CaspPI grouped with the PI/GLO lineages, and the C domain of CaspPI contained a conserved PI motif. CaspAP3 and CaspPI mRNA expression was mainly concentrated in petals and stamens but occurred at the primordium to mature stages of petals and stamens during morphological differentiation of organs. The expression of both factors peaked earlier in stamens than in petals, and the peak expression for both in petals appeared at the stage of rapid elongation. Meanwhile, the time of expression was the same with morphological differentiation of petals and stamens.

Key words: Catalpa speciosa, flower development, B-class MADS-box genes, gene cloning, the expression of gene

Danlong Jing, Yan Xia, Shougong Zhang, Junhui Wang. Expression Analysis of B-class MADS-box Genes from Catalpa speciosa[J]. Chinese Bulletin of Botany, 2016, 51(2): 210-217.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB15053

https://www.chinbullbotany.com/EN/Y2016/V51/I2/210

Figures/Tables 7

Table 1 The sequences of primers

Primer name	Primer sequences (5′→ 3′)
FLAP3F	GCAAACTCAAATCTTGAAAATC
FLAP3R	CACATTATGCGACAGAATTCAT
FLPIF	GAGAACCAAATTAAGAGAAAGACCA
FLPIR	GCACCAAGACAACCACATACGTA
RTactinF	ATGATGCTCCAAGAGCTGTG
RTactinR	AGCAAGATCGAGACGTAGGA
RTAP3F	TACATCAGCCCCATTATAACGA
RTAP3R	TTATTCAAGCAAGGCAAACGTG
RTPIF	ACCACAAGTTGTCTGGGAAGAG
RTPIR	AATCTGCAACTCCCTGATGATC
qRTactinF	GATGATGCTCCAAGAGCTGT
qRTactinR	TCCATATCATCCCAGTTGCT
qRTAP3F	CTTGAAGAAGCTGAAGGAGGTT
qRTAP3R	CTTGCTGGTATCAATCTGGTTG
qRTPIF	GAGAATGACAGCATGCAGATTG
qRTPIR	ATTATCGTCCATTGGATCCAGA

Table 2 The size of the flower buds of Catalpa speciosa

Length of flower buds (mm)	The stages and morphology of C. speciosa
1−3	In early stage of floral buds formation, the petals and stamens differentiated and elongated rapidly
4−13	At flower buds elongation stage, the size of petals and stamens increased rapidly
30−55	At flowering phase, the stamens reached mature stage, and the petals completely opened

Figure 1 Phylogenetic analysis of AP3 and PI subfamily

Figure 2 Comparison of deduced amino acid sequences encoded by CaspAP3 and CaspPI, and related members of AP3 and PI subfamily The first underlined regions represent the MADS domain and the second K domain, the region between the MADS- and K-domain is I domain. The PI-derived motif, euAP3 motif and PI motif are boxed.

Figure 3 Semi-quantitative RT-PCR of CaspAP3 and CaspPI gene in different tissues of Catalpa speciosa ste: Stem; j-le: Juvenile-leaf; m-le: Mature leaves; sep: Sepal; pet: Petal; sta: Stamen; car: Carpel; cap: Capsule; see: Seed

Figure 4 The characterization of the floral organ in Catalpa speciosa (A)−(C) The flower buds of 1−3 mm, the newly differentiated petal and stamen; (D)−(F) The flower buds of 4−13 mm, the sizes of petal and stamen increased rapidly; (G), (H) The flower buds of 30−55 mm, the petal and stamen reached mature stage, and the flowers reached anthesis. pe: Petal; sta: Stamen. Bar=500 μm

Figure 5 Quantitative real-time PCR analysis of CaspAP3 and CaspPI expressed in floral buds of Catalpa speciosa at different developmental stages (A) Relative expression patterns of CaspAP3 and CaspPI at differentiation stage of flowers; (B) Relative expression patterns of CaspAP3 and CaspPI at elongation and flowering stages of flowers

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