Identification of CibHLH1 and its effect on photosynthetic characteristics in Chrysanthemum indicum var. aromaticum

doi:10.11686/cyxb2023087

Abstract

Abstract:

The basic helix-loop-helix （bHLH） transcription factors are the second largest transcription factor family in plants and play an important role in plant growth and development， secondary metabolism and response to environmental stress. In this study， we cloned CibHLH1 using the leaf cDNA of Chrysanthemum indicum var. aromaticum as the template. The cloned gene has a 738 bp full-length sequence， and a 630 bp open reading frame. Multiple sequence comparison and phylogenetic investigation showed that CibHLH1 had the highest homology with CmbHLH1 of Chrysanthemum morifolium. The expression level of CibHLH1 was the highest in the flowers and lowest in roots. Subcellular localization revealed that the gene was located in the nucleus. CibHLH1 was transformed into tobacco （Nicotiana tabacum） by the leaf disk method to further study its functions. We found that the leaves of overexpressed tobacco became yellow and the contents of chlorophyll a， chlorophyll b and total chlorophyll were significantly reduced. The light response curve and photosynthetic physiological indexes of transgenic tobacco were evaluated， the results showed that overexpression of CibHLH1 decreased the maximum net photosynthetic rate， apparent quantum yield， light saturation point and water use efficiency， but increased the transpiration rate， stomatal conductance and light compensation point. In summary， CibHLH1 negatively affected the photosynthetic capacity of plants by reducing the content of photosynthetic pigments.

Key words: Chrysanthemum indicum var. aromaticum, CibHLH1, chlorophyll, photosynthetic characteristics

Jin-xiu HAN, Bin CHEN, Yan-ting LIU, Ru MENG, Li-yan JIN, Miao HE. Identification of CibHLH1 and its effect on photosynthetic characteristics in Chrysanthemum indicum var. aromaticum[J]. Acta Prataculturae Sinica, 2024, 33(1): 89-101.

Figures/Tables 12

References 57

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引物名称Primer name	引物序列Primer sequence （5′-3′）
bHLH-F	ATTTTACCAAACAGCGTCTTACGAGAAC
bHLH-R	TTAGGCAACCGGAGGGCGA
bHLH-1-F	AGTGCGGAACAAGATGGCTCA
bHLH-1-R	GGGCTTCCCAGGTTCAAGG
CmUBI-F	AGCTGAGCAGACTCCCGATG
CmUBI-R	AGGCGAATCATCAGTACCAAGT
CibHLH1-KpnI	GGTACCATGGTTTCACCGGAGACTACTACTAATT
CibHLH1-SpeI	ACTAGTGGCAACCGGAGGGCGAAG
pBI121-T-F	TCATTTCATTTGGAGAGAACAC
pBI121-T-R	TTGCCAAATGTTTGAACGATC

引物名称Primer name	引物序列Primer sequence （5′-3′）
bHLH-F	ATTTTACCAAACAGCGTCTTACGAGAAC
bHLH-R	TTAGGCAACCGGAGGGCGA
bHLH-1-F	AGTGCGGAACAAGATGGCTCA
bHLH-1-R	GGGCTTCCCAGGTTCAAGG
CmUBI-F	AGCTGAGCAGACTCCCGATG
CmUBI-R	AGGCGAATCATCAGTACCAAGT
CibHLH1-KpnI	GGTACCATGGTTTCACCGGAGACTACTACTAATT
CibHLH1-SpeI	ACTAGTGGCAACCGGAGGGCGAAG
pBI121-T-F	TCATTTCATTTGGAGAGAACAC
pBI121-T-R	TTGCCAAATGTTTGAACGATC

株系 Plants	最大净光合速率 Maximum net photosynthetic rate （μmol·m^-2·s^-1）	光补偿点 Light compensation point （μmol·m^-2·s^-1）	光饱和点 Light saturation point （μmol·m^-2·s^-1）	表观量子效率 Apparent photosynthetic quantum yield
WT	8.28±0.20a	12.11±6.56b	392.17±8.11a	0.0219±0.0014a
EV	8.35±0.65a	9.80±1.11b	363.50±13.52a	0.0241±0.0025a
B₁	5.13±0.51b	24.66±7.05a	325.85±49.52b	0.0174±0.0011b
B₂	4.98±0.88b	22.38±2.89a	306.69±25.21b	0.0176±0.0033b

株系 Plants	最大净光合速率 Maximum net photosynthetic rate （μmol·m^-2·s^-1）	光补偿点 Light compensation point （μmol·m^-2·s^-1）	光饱和点 Light saturation point （μmol·m^-2·s^-1）	表观量子效率 Apparent photosynthetic quantum yield
WT	8.28±0.20a	12.11±6.56b	392.17±8.11a	0.0219±0.0014a
EV	8.35±0.65a	9.80±1.11b	363.50±13.52a	0.0241±0.0025a
B₁	5.13±0.51b	24.66±7.05a	325.85±49.52b	0.0174±0.0011b
B₂	4.98±0.88b	22.38±2.89a	306.69±25.21b	0.0176±0.0033b

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