More Like this

1. Overexpression of two CDPKs from wild Chinese grapevine enhances powdery mildew resistance in Vitis vinifera and Arabidopsis

   https://doi.org/10.1111/nph.17285  

   Hu, Yang ; Cheng, Yuan ; Yu, Xuena ; Liu, Jie ; Yang, Lushan ; Gao, Yurong ; Ke, Guihua ; Zhou, Min ; Mu, Bo ; Xiao, Shunyuan ; et al ( March 2021 , New Phytologist)

   Calcium‐dependent protein kinases (CDPKs) play vital roles in metabolic regulations and stimuli responses in plants. However, little is known about their function in grapevine.

   Here, we report thatVpCDPK9andVpCDPK13, two paralogousCDPKsfromVitis pseudoreticulataaccession Baihe‐35‐1, appear to positively regulate powdery mildew resistance. The transcription of them in leaves of ‘Baihe‐35‐1’ were differentially induced upon powdery mildew infection. Overexpression ofVpCDPK9‐YFPorVpCDPK13‐YFPin theV. viniferasusceptible cultivar Thompson Seedless resulted in enhanced resistance to powdery mildew (YFP, yellow fluorescent protein). This might be due to elevation of SA and ethylene production, and excess accumulation of H₂O₂and callose in penetrated epidermal cells and/or the mesophyll cells underneath.

   Ectopic expression ofVpCDPK9‐YFPin Arabidopsis resulted in varied degrees of reduced stature, pre‐mature senescence and enhanced powdery mildew resistance. However, these phenotypes were abolished inVpCDPK9‐YFPtransgenic lines impaired in SA signaling (pad4sid2) or ethylene signaling (ein2). Moreover, both of VpCDPK9 and VpCDPK13 were found to interact with and potentially phosphorylate VpMAPK3, VpMAPK6, VpACS1 and VpACS2in vivo(ACS, 1‐aminocyclopropane‐1‐carboxylic acid (ACC) synthase; MAPK, mitogen‐activated protein kinase).

   These results suggest thatVpCDPK9andVpCDPK13contribute to powdery mildew resistance via positively regulating SA and ethylene signaling in grapevine.

    

   more » « less
2. Jasmonic acid and salicylic acid play minor roles in stomatal regulation by CO 2 , abscisic acid, darkness, vapor pressure deficit and ozone

   https://doi.org/10.1111/tpj.15430  

   Zamora, Olena ; Schulze, Sebastian ; Azoulay‐Shemer, Tamar ; Parik, Helen ; Unt, Jaanika ; Brosché, Mikael ; Schroeder, Julian I. ; Yarmolinsky, Dmitry ; Kollist, Hannes ( August 2021 , The Plant Journal)

   Jasmonic acid (JA) and salicylic acid (SA) regulate stomatal closure, preventing pathogen invasion into plants. However, to what extent abscisic acid (ABA), SA and JA interact, and what the roles of SA and JA are in stomatal responses to environmental cues, remains unclear. Here, by using intact plant gas‐exchange measurements in JA and SA single and double mutants, we show that stomatal responsiveness to CO₂, light intensity, ABA, high vapor pressure deficit and ozone either did not or, for some stimuli only, very slightly depended upon JA and SA biosynthesis and signaling mutants, includingdde2, sid2, coi1,jai1,myc2andnpr1alleles. Although the stomata in the mutants studied clearly responded to ABA, CO₂, light and ozone, ABA‐triggered stomatal closure innpr1‐1was slightly accelerated compared with the wild type. Stomatal reopening after ozone pulses was quicker in thecoi1‐16mutant than in the wild type. In intact Arabidopsis plants, spraying with methyl‐JA led to only a modest reduction in stomatal conductance 80 min after treatment, whereas ABA and CO₂induced pronounced stomatal closure within minutes. We could not document a reduction of stomatal conductance after spraying with SA. Coronatine‐induced stomatal opening was initiated slowly after 1.5–2.0 h, and reached a maximum by 3 h after spraying intact plants. Our results suggest that ABA, CO₂and light are major regulators of rapid guard cell signaling, whereas JA and SA could play only minor roles in the whole‐plant stomatal response to environmental cues in Arabidopsis andSolanum lycopersicum(tomato).

    

   more » « less
3. A role for calcium‐dependent protein kinases in differential CO 2 ‐ and ABA‐controlled stomatal closing and low CO 2 ‐induced stomatal opening in Arabidopsis

   https://doi.org/10.1111/nph.17079  

   Schulze, Sebastian ; Dubeaux, Guillaume ; Ceciliato, Paulo H. O. ; Munemasa, Shintaro ; Nuhkat, Maris ; Yarmolinsky, Dmitry ; Aguilar, Jaimee ; Diaz, Renee ; Azoulay‐Shemer, Tamar ; Steinhorst, Leonie ; et al ( December 2020 , New Phytologist)

   Low concentrations of CO₂cause stomatal opening, whereas [CO₂] elevation leads to stomatal closure. Classical studies have suggested a role for Ca²⁺and protein phosphorylation in CO₂‐induced stomatal closing. Calcium‐dependent protein kinases (CPKs) and calcineurin‐B‐like proteins (CBLs) can sense and translate cytosolic elevation of the second messenger Ca²⁺into specific phosphorylation events. However, Ca²⁺‐binding proteins that function in the stomatal CO₂response remain unknown.

   Time‐resolved stomatal conductance measurements using intact plants, and guard cell patch‐clamp experiments were performed.

   We isolatedcpkquintuple mutants and analyzed stomatal movements in response to CO₂, light and abscisic acid (ABA). Interestingly, we found thatcpk3/5/6/11/23quintuple mutant plants, but not other analyzedcpkquadruple/quintuple mutants, were defective in high CO₂‐induced stomatal closure and, unexpectedly, also in low CO₂‐induced stomatal opening. Furthermore, K⁺‐uptake‐channel activities were reduced incpk3/5/6/11/23quintuple mutants, in correlation with the stomatal opening phenotype. However, light‐mediated stomatal opening remained unaffected, and ABA responses showed slowing in some experiments. By contrast, CO₂‐regulated stomatal movement kinetics were not clearly affected in plasma membrane‐targetedcbl1/4/5/8/9quintuple mutant plants.

   Our findings describe combinatorialcpkmutants that function in CO₂control of stomatal movements and support the results of classical studies showing a role for Ca²⁺in this response.

    

   more » « less
4. Increased adaxial stomatal density is associated with greater mesophyll surface area exposed to intercellular air spaces and mesophyll conductance in diverse C 4 grasses

   https://doi.org/10.1111/nph.16106  

   Pathare, Varsha S. ; Koteyeva, Nuria ; Cousins, Asaph B. ( September 2019 , New Phytologist)

   Mesophyll conductance (g_m) is the diffusion ofCO₂from intercellular air spaces (IAS) to the first site of carboxylation in the mesophyll cells. In C₃species,g_mis influenced by diverse leaf structural and anatomical traits; however, little is known about traits affectingg_min C₄species.

   To address this knowledge gap, we used online oxygen isotope discrimination measurements to estimateg_mand microscopy techniques to measure leaf structural and anatomical traits potentially related tog_min 18 C₄grasses.

   In this study,g_mscaled positively with photosynthesis and intrinsic water‐use efficiency (TE_i), but not with stomatal conductance. Also,g_mwas not determined by a single trait but was positively correlated with adaxial stomatal densities (SD_ada), stomatal ratio (SR), mesophyll surface area exposed toIAS(S_mes) and leaf thickness. However,g_mwas not related to abaxial stomatal densities (SD_aba) and mesophyll cell wall thickness (T_CW).

   Our study suggests that greaterSD_adaandSRincreasedg_mby increasingS_mesand creating additional parallel pathways forCO₂diffusion inside mesophyll cells. Thus,SD_ada,SRandS_mesare important determinants of C₄‐g_mand could be the target traits selected or modified for achieving greaterg_mandTE_iin C₄species.

    

   more » « less
5. The Arabidopsis heterotrimeric G‐protein β subunit, AGB 1, is required for guard cell calcium sensing and calcium‐induced calcium release

   https://doi.org/10.1111/tpj.14318  

   Jeon, Byeong Wook ; Acharya, Biswa R. ; Assmann, Sarah M. ( April 2019 , The Plant Journal)

   Cytosolic calcium concentration ([Ca²⁺]_cyt) and heterotrimeric G‐proteins are universal eukaryotic signaling elements. In plant guard cells, extracellular calcium (Ca_o) is as strong a stimulus for stomatal closure as the phytohormone abscisic acid (ABA), but underlying mechanisms remain elusive. Here, we report that the sole Arabidopsis heterotrimeric Gβ subunit,AGB1, is required for four guard cell Ca_oresponses: induction of stomatal closure; inhibition of stomatal opening; [Ca²⁺]_cytoscillation; and inositol 1,4,5‐trisphosphate (InsP3) production. Stomata in wild‐type Arabidopsis (Col) and in mutants of the canonical Gα subunit,GPA1, showed inhibition of stomatal opening and promotion of stomatal closure by Ca_o. By contrast, stomatal movements ofagb1mutants andagb1/gpa1double‐mutants, as well as those of theagg1agg2 Gγ double‐mutant, were insensitive to Ca_o. These behaviors contrast withABA‐regulated stomatal movements, which involveGPA1 andAGB1/AGG3 dimers, illustrating differential partitioning of G‐protein subunits among stimuli with similar ultimate impacts, which may facilitate stimulus‐specific encoding.AGB1knockouts retained reactive oxygen species andNOproduction, but lostYC3.6‐detected [Ca²⁺]_cytoscillations in response to Ca_o, initiating only a single [Ca²⁺]_cytspike. Experimentally imposed [Ca²⁺]_cytoscillations restored stomatal closure inagb1. Yeast two‐hybrid and bimolecular complementation fluorescence experiments revealed thatAGB1 interacts with phospholipase Cs (PLCs), and Ca_oinduced InsP3 production in Col but not inagb1. In sum, G‐protein signaling viaAGB1/AGG1/AGG2 is essential for Ca_o‐regulation of stomatal apertures, and stomatal movements in response to Ca_oapparently require Ca²⁺‐induced Ca²⁺release that is likely dependent on Gβγ interaction withPLCs leading to InsP3 production.

    

   more » « less