Pulp A Different Class Zip !!LINK!!

I think it would be really helpful if .pdx.zip files were "first-class" citizens, and were supported by the simulator and playdate itself -- is this feature missing as a performance tradeoff? Sideloading through the website supports "zipped pdx files", but sideloading over USB doesn't. It means that players who download "pdx.zip" files via itch have to go through the technical step of unzipping the file.

Pulp using .zip instead of .pdx.zip was most likely a stylistic choice. It's also possible that downloaded games might (or might one day) contain other files alongside the .pdx, which would make the double extension inappropriate. But also, I don't think MacOS can be taught to treat .pdx.zip as a distinct file type from .zip, which is probably why we didn't try to solve this differently.

Schematic representation of HD-ZIP genes and their structural distribution split them into four classes (HD-ZIP I, HD-ZIP II, HD-ZIP III, and HD-ZIP IV). HD, LZ, START, SAD, and MEKHLA can be seen in the decoded form in the text.

There are nine genes in the Arabidopsis HD-ZIP II subfamily. The main role of this class in plant development is their shade-avoiding mechanism during the photosynthetic process [29,30,31]. For example, one member of class II subfamily ATHB2, when overexpressed in Arabidopsis, unfolded its role in plant development under illumination conditions [32]. On the other hand, microarray analysis revealed that HAT2, a member of the class II HD-ZIP gene family, was significantly influenced by the auxin during the seedlings stage [33]. To confirm that, Arabidopsis plants overexpressing the HAT2 gene produced epinastic cotyledons, long hypocotyls, long petioles, and small leaves. All these traits resembled to the mutants, generating auxin in high quantity [33,34]. Fruit ripening is an important qualitative factor that defines the fate market value of postharvest produces. Ethylene is generally considered a potent regulator of the fruit ripening process. In this regard, the overexpression of PpHB.G7, a class II HD-ZIP family gene in peach (Prunus persica), mediates the ripening process by altering the expression and production of ethylene biosynthesis genes and ethylene, respectively [35]. In a recent study, the rice (Oryza sativa) sgd2 gene was found responsible for small grain size and dwarf plant phenotype. The study further showed that the sgd2 gene is a transcriptional suppresser of GA biosynthetic genes, particularly suppressing the generation of endogenous GA1 [36]. The majority of the class II HD-ZIP genes that are differentially expressed in various plant tissues confer their importance in regulating plant developmental activities.

Arabidopsis genome contains five members of the class III HD-ZIP gene family. Numerous mutants of these genes have been reported previously. Most of the class III genes are responsible for sustaining the normal organ polarity and shoot apical meristem (SAM) [37]. Single loss of HD-ZIP III protein function does not display any obvious phenotypic changes. However, a double or triple mutant of class III genes such as phb-6/phv-5/rev-9 lacked SAM along with single abaxialized cotyledon, suggesting their overlapping nature [38]. Additionally, overexpression of Arabidopsis ATHB8 hastened the xylem formation because of the ectopic production of procambial cells [39]. In contrast, loss of function of ATHB8 failed to show any physiological and morphological changes [39]. The ATHB15 gained the icu4-1 function allele, resulting in an abnormal arrangement of root meristem and more number of lateral roots production than the wild type (WT) [40]. Taken together, the aforementioned statements elucidated the crucial role of class III genes in root formation and vascular development. Another study [41] supported the notion by reporting the role of class III gene in nodule formation, root development, and vascular activities regulation. The results highlighted that GmHD-ZIP III 2 demonstrated strong interaction with GmZPR3d, ensuing in the ectopic formation of secondary root xylem and also a dominant expression of soybean (Glycine max) vessel-specific genes [41].

Alteration in the leaf mesophyll anatomy by genetic modification is potentially a promising tool for improving the physiological functions of trees by improving leaf photosynthesis. Homeodomain leucine zipper (HD-Zip) transcription factors are candidates for anatomical alterations of leaves through modification of cell multiplication, differentiation, and expansion. Full-length cDNA encoding a Eucalyptus camaldulensis HD-Zip class II transcription factor (EcHB1) was over-expressed in vivo in the hybrid Eucalyptus GUT5 generated from Eucalyptus grandis and Eucalyptus urophylla. Overexpression of EcHB1 induced significant modification in the mesophyll anatomy of Eucalyptus with enhancements in the number of cells and chloroplasts on a leaf-area basis. The leaf-area-based photosynthesis of Eucalyptus was improved in the EcHB1-overexpression lines, which was due to both enhanced CO2 diffusion into chloroplasts and increased photosynthetic biochemical functions through increased number of chloroplasts per unit leaf area. Additionally, overexpression of EcHB1 suppressed defoliation and thus improved the growth of Eucalyptus trees under drought stress, which was a result of reduced water loss from trees due to the reduction in leaf area with no changes in stomatal morphology. These results gave us new insights into the role of the HD-Zip II gene.

Leaf photosynthetic functions are key traits for the determination of tree growth. Enhancement of leaf photosynthesis will thus be a possible measure for improvement in the productivity of forest trees. Eucalyptus species are one of the most widely-planted multipurpose tree species in the world1,2, which are used as a source of paper pulp, wood, and timber. Eucalyptus species have a high growth rate and high photosynthetic rate, while soil water availability strongly limits their growth3,4; therefore, improvement in leaf photosynthesis and tree growth, as well as raising drought tolerance under limited water resources, are important issues for improving the production of Eucalyptus trees.

One of the promising measures to improve leaf photosynthesis is to modify the leaf mesophyll anatomy. Leaf photosynthesis is limited by (1) CO2 diffusion through the stomata, (2) CO2 diffusion through the mesophyll, and (3) photosynthetic biochemistry, in which both CO2 diffusion through the mesophyll and photosynthetic biochemistry are strongly affected by the leaf mesophyll anatomy, in particular the number of chloroplasts5,6,7,8. Thick and dense leaf mesophyll tissue generally has a high number of chloroplasts and thus involves high photosynthetic functions through high mesophyll CO2 diffusion and a large amount of carboxylation enzymes within a single species9,10,11 and among different species of angiosperms and ferns8. Furthermore, the above alterations in the leaf mesophyll anatomy also potentially improve both the growth and drought tolerance of tree leaves through improvement in the water-use efficiency, when an increase in water-use efficiency is not accompanied by a concurrent decline in CO2 fixation limited by mesophyll or photosynthetic biochemistry12. Although a high photosynthetic rate generally corresponds to high water loss from the stomata due to large stomatal conductance and high hydraulic conductance among different species13, an increase in the photosynthetic rate with little change in stomatal conductance has been reported in Eucalyptus leaves overexpressing radish aquaporin RsPIP2;114.

Significant enhancements in the area-based leaf photosynthetic functions were obtained in Eucalyptus trees overexpressing EcHB1 (Fig. 3). This is the first study showing the effect of an HD-Zip class II transcription factor on the leaf photosynthetic function. Leaf photosynthesis is limited by 1) biochemical activities and 2) CO2 diffusion through the stomata and mesophyll6,7,8. Overexpression of EcHB1 in the leaf induced significant alterations in the leaf mesophyll anatomy (Fig. 2, Table 1), which contributed to significant enhancements in the area-based leaf photosynthetic functions (Fig. 3) through enhancements in both the biochemical activities (Vcmax and J, Fig. 3) and CO2 diffusion through the mesophyll (gm, Fig. 4). The principle component analysis of leaf anatomical traits indicated that the marked increase in the mesophyll fraction (1.5-1.9-fold, Fig. 2B) was mainly due to the increase in the number of mesophyll cells (1.4-fold, Fig. 5B, Table 1). The increase in the number of mesophyll cells is related to an increase in the number of chloroplasts (1.5-fold, Fig. 5B, Table 1), which may induce enhancements in the photosynthetic biochemistry, e.g., the carboxylation enzyme Rubisco that regulates the velocity of carboxylation (Vcmax) and photochemical efficiency and the Calvin cycle that regulates the electron transport rate (J, Fig. 3B). When the relationship between the leaf anatomical traits and biochemical parameters was analyzed, a high Vcmax corresponded to a large number of chloroplasts in angiosperms and ferns8,23.

Overexpression of EcHB1 induced marked enhancement in the mesophyll diffusional conductance, gm (1.8-1.9-fold, Fig. 4A), which is strongly affected by alterations in the mesophyll anatomy (Fig. 5A); the significant increase in the number of chloroplasts (1.5-fold) with a decrease in chloroplast size (Table 1) resulted in a 1.6-fold increase in the chloroplast surface area (Sc, Table 1) for CO2 diffusion5. The concurrent increases in Sc and gm in the EcHB1-overexpressed lines (Fig. 4C), together with the concurrent increases in gm and A (Fig. 4C), supports the significant role of Sc for determining gm, and thus, photosynthesis. On the other hand, overexpression of EcHB1 induced little alteration in the stomatal conductance, gs (Fig. 4A). The stomatal density and size strongly affect stomatal conductance, gs24. Low stomatal density as well as large stomatal size reduces gs, because low stomatal density reduces the total stomatal pore area in leaves, while large stomatal size increases the diffusion path length25. The small change in stomatal conductance (gs) by the overexpression of EcHB1 (Fig. 4A) may be due to the lack of alterations in both the stomatal density and size (Table 1). CO2 diffusion through the stomata and mesophyll imposes almost comparable limitations in photosynthesis in different tree groups including deciduous, evergreens, and conifers7. The present study for Eucalyptus trees showed that the mesophyll tended to impose lower limitations in the overexpression lines EcHB1-2 and EcHB1-10 compared to GUT5 (Fig. 4B). This result reflects the enhancement of gm by the overexpression of EcHB1 (Fig. 4A). 2b1af7f3a8