Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (2): 193-202.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0506

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Responses of Choloroplast Specific Protein Profile to Different Stomatal Densities in Nicotiana benthamiana

SA Shi-juan1,2(), WU Han-yu1,2, WEN Yuan1,2, CHEN Xue-na1,2, ZHENG Rui1,2, YAO Xin-ling1,2()   

  1. 1. Key Laboratory of Modern Molecular Breeding for Dominant and Special Crops in Ningxia, Ningxia University, Yinchuan 750021
    2. Education Ministry Lab for Protection and Utilization on Bioresource in the West China, Ningxia University, Yinchuan 750021
  • Received:2022-04-24 Online:2023-02-26 Published:2023-03-07

Abstract:

CO2 enters chloroplast through stomata. How chloroplast is in response to stomatal density in molecular level is unknown yet. In this study, transgenic Nicotiana benthamiana lines, showing either significantly higher(HSD)or lower(LSD)leaf stomatal density, respectively, were generated through regulation of NtEPF 2(Nicotiana benthamiana EPIDERMAL PATTERNING FACTOR 2)expression in vivo. Transgenic lines were characterized with chlorophyll accumulation, photosynthetic parameters and leaf chloroplast differential expression proteins(DEPs)based on iTRAQ(isobaric tags for relative and absolute quantitation). The response of chloroplast composition to stomatal density variation was expected to be revealed. The results showed that accumulation of DEPs, metabolizing PSI-PSII-LHCII complex formation and accelerating its electron transfer were up-regulated significantly, following down-regulation of DEPs for PSI photoprotection and light damage repair in HSD lines. Therefore, chlorophyll accumulation and net photosynthetic rate were 43% and 67% higher in HSD lines than control. On the other hand, attributing to strong photophosphorylation ATP biosynthesis at the early stage of leaf development, chlorophyll accumulation and photosynthetic parameter in LSD lines were significantly higher than in control. Chlorophyll accumulation went down along with growing leaf age. The result indicated that the increase in stomatal density accelerated PSI-PSII-LHCII complex formation and its electron transfer, and reduced photoprotection. Thus, chlorophyll accumulations in leaf at all ages were maintained at a higher level. Reversely, the decrease in stomatal density resulted in an upside-down mode for chlorophyll accumulations along with leaf ages owing to unbalance between ATP biosynthesis and carbon fixation. The result not only contributes to further understanding how stomata work, also broadens our view to rise carbon fixation through stomata regulation.

Key words: iTRAQ, NtEPF 2, protein, chlorophyll, chloroplast, stomatal density, Nicotiana benthamiana