Nitric oxide modulates the influx of extracellular Ca2+ and actin filament organization during cell wall construction in Pinus bungeana pollen tubes.
Nitric oxide (NO) plays a key role in many physiological processes in plants, including pollen tube growth. Here, effects of NO on extracellular Ca(2+) flux and microfilaments during cell wall construction in Pinus bungeana pollen tubes were investigated. Extracellular Ca(2+) influx, the intracellular Ca(2+) gradient, patterns of actin organization, vesicle trafficking and cell wall deposition upon treatment with the NO donor S-nitroso-N-acetylpenicillamine (SNAP), the NO synthase (NOS) inhibitor N(omega)-nitro-L-arginine (L-NNA) or the NO scavenger 2-(4-carboxyphenyl)-4, 4, 5, 5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) were analyzed. SNAP enhanced pollen tube growth in a dose-dependent manner, while L-NNA and cPTIO inhibited NO production and arrested pollen tube growth. Noninvasive detection and microinjection of a Ca(2+) indicator revealed that SNAP promoted extracellular Ca(2+) influx and increased the steepness of the tip-focused Ca(2+) gradient, while cPTIO and L-NNA had the opposite effect. Fluorescence labeling indicated that SNAP, cPTIO and L-NNA altered actin organization, which subsequently affected vesicle trafficking. Finally, the configuration and/or distribution of cell wall components such as pectins and callose were significantly altered in response to L-NNA. Fourier transform infrared (FTIR) microspectroscopy confirmed the changes in the chemical composition of walls. Our results indicate that NO affects the configuration and distribution of cell wall components in pollen tubes by altering extracellular Ca(2+) influx and F-actin organization.
Wang
Yuhua
Y
Key Laboratory of Photosynthesis and Molecular Environmental Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.|Graduate School of Chinese Academy of Sciences, Beijing 100049, China.|The College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China.
Chen
Tong
T
Key Laboratory of Photosynthesis and Molecular Environmental Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
Zhang
Chunyang
C
Key Laboratory of Photosynthesis and Molecular Environmental Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.|Graduate School of Chinese Academy of Sciences, Beijing 100049, China.
Hao
Huaiqing
H
Key Laboratory of Photosynthesis and Molecular Environmental Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
Liu
Peng
P
Key Laboratory of Photosynthesis and Molecular Environmental Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.|Graduate School of Chinese Academy of Sciences, Beijing 100049, China.
Zheng
Maozhong
M
Key Laboratory of Photosynthesis and Molecular Environmental Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.|Graduate School of Chinese Academy of Sciences, Beijing 100049, China.
Baluška
František
F
Institute of Cellular and Molecular Botany, Rheinische Friedrich-Wilhelms-University Bonn, Department of Plant Cell Biology, Kirschallee 1, D-53115 Bonn, Germany.
Šamaj
Jozef
J
Institute of Cellular and Molecular Botany, Rheinische Friedrich-Wilhelms-University Bonn, Department of Plant Cell Biology, Kirschallee 1, D-53115 Bonn, Germany.|Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences, Akademicka 2, SK-95007, Nitra, Slovak Republic.|Palacky University Olomouc, Faculty of Natural Science, Olomouc 771 46, Czech Republic.
Lin
Jinxing
J
Key Laboratory of Photosynthesis and Molecular Environmental Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D001565:Benzoates / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002473:Cell Wall / Q000187:drug effects / Q000378:metabolism*; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism*; D018525:Germination / Q000187:drug effects; D005936:Glucans / Q000378:metabolism; D007093:Imidazoles / Q000494:pharmacology; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D008954:Models, Biological; D009569:Nitric Oxide / Q000096:biosynthesis / Q000494:pharmacology*; D019335:Nitroarginine / Q000494:pharmacology; D010368:Pectins / Q000378:metabolism; D028223:Pinus / Q000187:drug effects / Q000378:metabolism*; D053205:Pollen Tube / Q000166:cytology / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D058105:Polymerization / Q000187:drug effects; D011726:Pyridinium Compounds / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D026423:S-Nitroso-N-Acetylpenicillamine / Q000494:pharmacology; D017550:Spectroscopy, Fourier Transform Infrared; D013194:Staining and Labeling; D013997:Time Factors
D001565:Benzoates; C092350:FM 4-64; D005936:Glucans; D007093:Imidazoles; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; C079393:1,3-dihydroxy-4,4,5,5-tetramethyl-2-(4-carboxyphenyl)tetrahydroimidazole; D019335:Nitroarginine; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine; D010368:Pectins; C048306:callose; D002118:Calcium