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Tuska earman1/3/2024 Curre ntly, it is t heorize d 42 that the te mporal se paration o f CO 2 fixation is under cont rol of the circad ian clock 6,7 and that the 43 inverted stomatal c onductance c ould be a result of a change in clock regulation 8. 39 These trai ts coupled wit h the global i ssue of increas ed frequency and intensity of dr ought 2,3 have 40 generated an increa se in CAM researc h with the goa l of engineer ing these t raits in to C 3 plants, to 41 enable better drought res ponses and/ or improved drou ght toleranc e 4,5. ![]() CA M plants 37 exhibit im prove d photosy nthetic ef ficiency due to a te mporal se paration o f CO 2 fixation and 38 imp roved water-us e efficienc y due to inverted s tomatal conduc tance, relat ive to C 3 plants 1. 35 Plants that p hotosynthe size thro ugh crassula cean acid m etabolism ( CAM) are great 36 example s of how plants sync hronize biolog ical process es to the ir environm ent. The circad ian clock th us enables p lants to ac tivate the ap propriat e 34 processes in response to these re peating variables. Numerous env ironmental signals and stressors to plants are cy clic, such a s light ava ilability, 33 temperatu re, and predation. fedtschenkoi, fa cilitati ng the engin eering of CAM ma chinery 27 into non- CAM plan ts for sus tainabl e crop pro duction i n water- limited env ironme nts. Our resu lts provide n ew insights into th e mechanism of ci rcadian 26 control o f CAM-re lated ge nes in K. fedtsche nkoi cor e clock g enes an d construc ted a su bsequent 25 gene regulat ory network. W e also used L EM to ide ntify 24 stomata-related gene target s for K. We utili zed a new metri c 21 f or identif ying can didate cor e gene s of a perio dic gene ne twork an d then a pplied the Local Edge 22 Machine (L EM) algor ithm to infe r regul atory re lationshi ps betw een the ca ndidate co re clock 23 genes and orth ologs of known core c lock genes in K. The refore, we set out to inves tigate whe ther 19 core circa dian el ements in CA M plant s were re-phased d uring ev olution, o r whethe r netwo rks of 20 phase-sp ecific gen es were sim ply connec ted to di fferent c ore element s. The molecu lar mecha nisms behi nd how the cir cadian cloc k enabled th ese 18 physiol ogical diffe rences is not w ell understo od. C rassulacean ac id 15 metabolis m (CAM) photosynt hetic plant s represent an interesti ng case of circa dian regulati on of 16 gene e xpression as CO 2 fixatio n and sto matal mov ement in CA M plants display strong cir cadian 17 dynamics. ![]() The circ adian clock d rives tim e-specifi c gene expres sion, allo wing for as sociated 14 biologica l processes t o be active during c ertain tim es of the 24 h day. fedtschenkoi, facilitating the engineering of CAM machinery into non-CAM plants for sustainable crop production in water-limited environments. ![]() ![]() Our results provide new insights into the mechanism of circadian control of CAM-related genes in K. fedtschenkoi core clock genes and constructed a subsequent gene regulatory network. We also used LEM to identify stomata-related gene targets for K. We utilized a new metric for identifying candidate core genes of a periodic gene network and then applied the Local Edge Machine (LEM) algorithm to infer regulatory relationships between the candidate core clock genes and orthologs of known core clock genes in K. Therefore, we set out to investigate whether core circadian elements in CAM plants were re-phased during evolution, or whether networks of phase-specific genes were simply connected to different core elements. The molecular mechanisms behind how the circadian clock enabled these physiological differences is not well understood. Crassulacean acid metabolism (CAM) photosynthetic plants represent an interesting case of circadian regulation of gene expression as CO2 fixation and stomatal movement in CAM plants display strong circadian dynamics. The circadian clock drives time-specific gene expression, allowing for associated biological processes to be active during certain times of the 24 h day.
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