How to make lunar soil suitable for cultivation? - A review.

The investigation of lunar soil encompasses extensive periods, employs many improvement methods, and has generated several simulants. The improvement of lunar soil has recently garnered growing interest as an aspect of In-Situ Resource Utilization (ISRU) for regolith. It is crucial to clarify the challenges of utilizing lunar soil as a planting substrate to develop more effective techniques. This review presents a comprehensive analysis of research on improving lunar soil properties, highlights the disparities in mineral composition between real lunar soil (also called regolith) and simulated lunar soil, then details their deficiencies as planting substrates. Following an investigation of existing improvement methods, a dilemma of metals、salt precipitation and high pH caused by adding organic matter alone was noted, while the function of microbes (bacteria, algae, and lichens) in improvement processes was assessed. Finally, we present a perspective on future the lunar soil plantable research development based on the Bioregenerative Life Support System (BLSS). This review aims to promote the engineering application of lunar soil improvements and sustainable development. We hope that one day, regolith will enable plants to flourish on the Moon.

Identification of Apple Flower Development-Related Gene Families and Analysis of Transcriptional Regulation.

Apple (Malus domestica Borkh.) stands out as a globally significant fruit tree with considerable economic importance. Nonetheless, the orchard production of 'Fuji' apples faces significant challenges, including delayed flowering in young trees and inconsistent annual yields in mature trees, ultimately resulting in suboptimal fruit yield due to insufficient flower bud formation. Flower development represents a pivotal process influencing plant adaptation to environmental conditions and is a crucial determinant of successful plant reproduction. The three gene or transcription factor (TF) families, C2H2, DELLA, and FKF1, have emerged as key regulators in plant flowering regulation; however, understanding their roles during apple flowering remains limited. Consequently, this study identified 24 MdC2H2, 6 MdDELLA, and 6 MdFKF1 genes in the apple genome with high confidence. Through phylogenetic analyses, the genes within each family were categorized into three distinct subgroups, with all facets of protein physicochemical properties and conserved motifs contingent upon subgroup classification. Repetitive events between these three gene families within the apple genome were elucidated via collinearity analysis. qRT-PCR analysis was conducted and revealed significant expression differences among MdC2H2-18, MdDELLA1, and MdFKF1-4 during apple bud development. Furthermore, yeast two-hybrid analysis unveiled an interaction between MdC2H2-18 and MdDELLA1. The genome-wide identification of the C2H2, DELLA, and FKF1 gene families in apples has shed light on the molecular mechanisms underlying apple flower bud development.

Apple MdZAT5 mediates root development under drought stress.

Root plays an important role in plant drought tolerance, especially in horticultural crops like apples. However, the crucial regulator and molecular mechanism in root development of apple trees under drought are not well unknown. Cys2/His2-type Zinc-finger proteins are essential for plant response to drought, while the members of C2H2 Zinc-finger proteins in apple are largely unknown. In this study, we identified the members of the C1-2i subclass family of C2H2 Zinc-finger proteins in apple (Malus × domestica). Among them, MdZAT5 is significantly induced in apple roots under drought conditions and positively regulates apple root development under drought. Further investigation revealed that MdZAT5 positively regulates root development and root hydraulic conductivity by mediating the transcription level of MdMYB88 under drought stress. Taken together, our results demonstrate the importance of MdZAT5 in root development under drought in apple trees. This finding provides a new candidate direction for apple breeding for drought resistance.

The apple Ca2+/H+ exchanger MdCAX2L-2 functions positively in modulation of Ba2+ tolerance.

Calcium is an essential element for plant growth and development, and it plays an important role in the responses of plants to abiotic stress. High concentrations of heavy metal ions in soil significantly affect the yield and quality of crops and pose human health threats when these ions accumulate in edible organs. The Ca2+/H+ exchanger (CAX) family is a class of transporters that mediate the transmembrane transport of both Ca2+ and metal ions, and they are widely involved in regulating plant growth and development and stress responses. Here, we cloned an AtCAX2 ortholog, MdCAX2L-2, from apple. It is constitutively expressed in various apple tissues and significantly induced by Ca2+ and Ba2+ treatments. The MdCAX2L-2 protein is located in the vacuolar membrane in both plant and yeast cells. Overexpression of MdCAX2L-2 enhanced the tolerance of the yeast mutant K667 to high concentrations of Ca2+ and Ba2+. In addition, the role of MdCAX2L-2 in modulating Ba2+ tolerance was identified using MdCAX2L-2-overexpressing transgenic Arabidopsis plants and apple calli. Comparison of growth phenotypes and stress-related physiological indexes under BaCl2 treatment indicated that MdCAX2L-2 could enhance the Ba2+ tolerance of plants by promoting Ba2+ compartmentalization into the vacuoles and eliminating excess ROS. Our results provide insights that will aid future studies examining the function of CAX proteins in regulating stress tolerance in fruit crops, as well as their underlying mechanisms.

Si-enriched biochars improved soil properties, reduced Cd bioavailability while enhanced Cd translocation to grains of rice.

Biochar and silicon (Si) have been widely considered to play an important role in mitigating cadmium (Cd) toxicity. In this study, wild-type rice (WT, high-Si) and Si-deficient mutant rice (lsi1, low-Si) were used as raw materials to prepare biochar at 500℃; the Si concentrations of high- and low-Si biochar were 15.9% and 5.3%, respectively. The impacts of different application rates (0%, 2%, 4%) of high- and low-Si biochars on soil chemical properties, Si and Cd fractions and availability, Cd absorption, and translocation were investigated. The results showed that both types of biochars increased soil pH, soil available nitrogen, and available phosphorus and potassium; and promoted Si uptake and plant growth of rice. Soil available Si, CaCl2-Si, acetic-Si, H2O2-Si, oxalate-Si, and Na2CO3-Si were also increased by biochar supply, especially for high-Si biochar treatments. In addition, both types of biochars had no effects on soil total Cd, but reduced soil available Cd by 2-17% in early season 2022, and reduced oxidizable Cd and residual Cd. Biochar application did not influence Cd concentrations in roots, stems, and leaves, but significantly increased Cd uptake and transport from stems and leaves to grains. The results suggested that Si-rich biochar could improve soil nutrients, change soil Si/Cd fractions and availability, promote rice growth but increase the risk of Cd toxicity in grains, indicating the complex of straw biochar in remediating Cd-contaminated paddy soil.

MdNAC104 positively regulates apple cold tolerance via CBF-dependent and CBF-independent pathways.

Low temperature is the main environmental factor affecting the yield, quality and geographical distribution of crops, which significantly restricts development of the fruit industry. The NAC (NAM, ATAF1/2 and CUC2) transcription factor (TF) family is involved in regulating plant cold tolerance, but the mechanisms underlying these regulatory processes remain unclear. Here, the NAC TF MdNAC104 played a positive role in modulating apple cold tolerance. Under cold stress, MdNAC104-overexpressing transgenic plants exhibited less ion leakage and lower ROS (reactive oxygen species) accumulation, but higher contents of osmoregulatory substances and activities of antioxidant enzymes. Transcriptional regulation analysis showed that MdNAC104 directly bound to the MdCBF1 and MdCBF3 promoters to promote expression. In addition, based on combined transcriptomic and metabolomic analyses, as well as promoter binding and transcriptional regulation analyses, we found that MdNAC104 stimulated the accumulation of anthocyanin under cold conditions by upregulating the expression of anthocyanin synthesis-related genes, including MdCHS-b, MdCHI-a, MdF3H-a and MdANS-b, and increased the activities of the antioxidant enzymes by promoting the expression of the antioxidant enzyme-encoding genes MdFSD2 and MdPRXR1.1. In conclusion, this study revealed the MdNAC104 regulatory mechanism of cold tolerance in apple via CBF-dependent and CBF-independent pathways.

The cold-stress responsive gene DREB1A involved in low-temperature tolerance in Xinjiang wild walnut.

Background: Low-temperatures have the potential to be a serious problem for plants and can negatively affect the normal growth and development of walnuts. DREB1/CBF (Dehydration Responsive Element Binding Protein 1/C-repeat Binding Factor), one of the most direct transcription factors in response to low-temperature stress, may improve the resistance of plants to low-temperatures by regulating their functional genes. However, few studies have been conducted in walnut. The Xinjiang wild walnut is a rare wild plant found in China, with a large number of excellent trait genes, and is hardier than cultivated walnuts in Xinjiang. Methods: In this work, we identified all of the DREB1 members from the walnut genome and analyzed their expression levels in different tissues and during low-temperature stress on the Xinjiang wild walnut. The JfDREB1A gene of the Xinjiang wild walnut was cloned and transformed into Arabidopsis thaliana for functional verification. Results: There were five DREB1 transcription factors in the walnut genome. Among them, the relative expression level of the DREB1A gene was significantly higher than other members in the different tissues (root, stem, leaf) and was immediately un-regulated under low-temperature stress. The overexpression of the JfDREB1A gene increased the survival rates of transgenic Arabidopsis lines, mainly through maintaining the stability of cell membrane, decreasing the electrical conductivity and increasing the activities of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). Additionally, the expression levels of cold-inducible genes like AtKIN1, AtERD10, AtRD29A, AtCOR15A and AtCOR47, were significantly increased. These results showed that the JfDREB1A gene may play an important role in the response to cold stress of the Xinjiang wild walnut. This study contributes to our understanding of the molecular mechanism of the Xinjiang wild walnut's response to low-temperature stress and will be beneficial for developing walnut cultivars with improved cold resistance.

Characterization of soil microbial community activity and structure for reducing available Cd by rice straw biochar and Bacillus cereus RC-1.

The combination of biochar and specific bacteria has been widely applied to remediate Cadmium-contaminated soil. But little is known about how such composites affect the dynamic distribution of metal fractions. This process is accompanied by the alternations of soil properties and microbial community structures. Composite of rice straw biochar and Bacillus cereus RC-1 were applied to investigate its impacts on Cd alleviation and soil microbial diversity and structure. The bacterial/biochar composite treatment decreased the fraction of HOAc-extractable Cd by 38.82%, and increased residual Cd by 23.95% compared to the untreated control. Moreover, compared with the untreated control, the composite treatment significantly increased the soil pH by about 1.5 units, and the activities of catalase, urease and invertase enzymes were increased by 42.39%, 30.50% and 31.20%, respectively. Composite treatment increased soil bacterial and fungal alpha diversity, the relative abundance of Bacillus, Streptomyces, Arthrobacter, and Aspergillus species were also increased. Mantel test and correlation analysis indicated that the effects associated with fungal communities in influencing soil properties were lower than that those of bacterial communities by different treatment. Aggregated boosted tree (ABT) models analysis showed that soil chemical proprieties (as determined by SOM, CEC, AN, etc.,) contributed over 50% of the changes in bacterial and fungal communities by the composite treatment. The co-occurrence network results showed that all treatments enhanced the correlation between OUT groups and improved the possible relationships in the bacterial and fungal communities, especially the interrelationships between bacteria and fungi after the Cd fractions stabilized. These findings provide a new insight to optimal strategies for the remediation of Cd-contaminated soil.

Potential roles of melatonin and ABA on apple dwarfing in semi-arid area of Xinjiang China.

Dwarfing is a typic breeding trait for mechanical strengthening and relatively high yield in modern apple orchards. Clarification of the mechanisms associated with dwarfing is important for use of molecular technology to breed apple. Herein, we identified four dwarfing apple germplasms in semi-arid area of Xinjiang, China. The internodal distance of these four germplasms were significantly shorter than non-dwarfing control. Their high melatonin (MT) contents are negatively associated with their malondialdehyde (MDA) levels and oxidative damage. In addition, among the detected hormones including auxin (IAA), gibberellin (GA), brassinolide (BR), zeatin-riboside (ZR), and abscisic acid (ABA), only ABA and ZR levels were in good correlation with the dwarfing phenotype. The qPCR results showed that the expression of melatonin synthetic enzyme genes MdASMT1 and MdSNAT5, ABA synthetic enzyme gene MdAAO3 and degradative gene MdCYP707A, ZR synthetic enzyme gene MdIPT5 all correlated well with the enhanced levels of MT, ABA and the reduced level of of ZR in the dwarfing germplasms. Furthermore, the significantly higher expression of ABA marker genes (MdRD22 and MdRD29) and the lower expression of ZR marker genes (MdRR1 and MdRR2) in all the four dwarf germplasms were consistent with the ABA and ZR levels. Considering the yearly long-term drought occurring in Xinjiang, China, it seems that dwarfing with high contents of MT and ABA may be a good strategy for these germplasms to survive against drought stress. This trait of dwarfing may also benefit apple production and breeding in this semi-arid area.

Effect of rice straw biochar on three different levels of Cd-contaminated soils: Cd availability, soil properties, and microbial communities.

Biochar can be effective in immobilizing soil cadmium (Cd), but the difference in its immobilization mechanisms for different levels of Cd-contaminated soils was overlooked. In this study, rice straw biochar (BC) was added to three Cd-contaminated soils following 180 days of incubation, in the process of which the dynamic changes of Cd speciation, soil properties and microbial community diversity were determined. BC could significantly reduce the ratio of acid-soluble in the three soils, especially in light and medium Cd-contaminated soils by more than 20%. The addition of biochar could significantly increase the soil pH, soil organic matter, cation exchange capacity, and the activities of catalase, but decrease the richness and diversity of bacterial communities in all soils. The associations between microbial communities were inhibited in light and medium Cd-contaminated soils, but promoted in heavy Cd-contaminated soils. Furthermore, the main pathway of BC effect on soil Cd availability was also analyzed by partial least squares model (PLS-PM), which indicated that BC indirectly reduced Cd availability mainly by regulating the microbial community in light Cd-contaminated soil, whereas BC directly reduced Cd availability primarily by its own adsorption in medium and heavy Cd-contaminated soils. This research deepened understanding of the mechanisms of stabilization of Cd by biochar for agricultural soils.

Transcriptome profiling based on Illumina- and SMRT-based RNA-seq reveals circadian regulation of key pathways in flower bud development in walnut.

Flower bud development is a defining feature of walnut, which contributes to the kernel yield, yield stability, fruit quality and commodity value. However, little is known about the mechanism of the flower bud development in walnut. Here, the stages of walnut female flower bud development were divided into five period (P01-05) by using histological observation. They were further studied through PacBio Iso-Seq and RNA-seq analysis. Accordingly, we obtained 52,875 full-length transcripts, where 4,579 were new transcripts, 3,065 were novel genes, 1,437 were consensus lncRNAs and 20,813 were alternatively spliced isoforms. These transcripts greatly improved the current genome annotation and enhanced our understanding of the walnut transcriptome. Next, RNA sequencing of female flower buds at five periods revealed that circadian rhythm-plant was commonly enriched along with the flower bud developmental gradient. A total of 14 differentially expressed genes (DEGs) were identified, and six of them were confirmed by real-time quantitative analysis. Additionally, six and two differentially expressed clock genes were detected to be regulated by AS events and lncRNAs, respectively. All these detected plant circadian genes form a complex interconnected network to regulate the flower bud development. Thus, investigation of key genes associated with the circadian clock could clarify the process of flower bud development in walnut.

Integrated transcriptomic and proteomic analysis reveals the complex molecular mechanisms underlying stone cell formation in Korla pear.

Korla pear (Pyrus sinkiangensis Yü) is a landrace selected from a hybrid pear species in the Xinjiang Autonomous Region in China. In recent years, pericarp roughening has been one of the major factors that adversely affects fruit quality. Compared with regular fruits, rough-skin fruits have a greater stone cell content. Stone cells compose sclerenchyma tissue that is formed by secondary thickening of parenchyma cell walls. In this work, we determined the main components of stone cells by isolating them from the pulp of rough-skin fruits at the ripening stage. Stone cell staining and apoptosis detection were then performed on fruit samples that were collected at three different developmental stages (20, 50 and 80 days after flowering (DAF)) representing the prime, late and stationary stages of stone cell differentiation, respectively. The same batches of samples were used for parallel transcriptomic and proteomic analysis to identify candidate genes and proteins that are related to SCW biogenesis in Korla pear fruits. The results showed that stone cells are mainly composed of cellulose (52%), hemicellulose (23%), lignin (20%) and a small amount of polysaccharides (3%). The periods of stone cell differentiation and cell apoptosis were synchronous and primarily occurred from 0 to 50 DAF. The stone cell components increased abundantly at 20 DAF but then decreased gradually. A total of 24,268 differentially expressed genes (DEGs) and 1011 differentially accumulated proteins (DAPs) were identified from the transcriptomic and proteomic data, respectively. We screened the DEGs and DAPs that were enriched in SCW-related pathways, including those associated with lignin biosynthesis (94 DEGs and 31 DAPs), cellulose and xylan biosynthesis (46 DEGs and 18 DAPs), S-adenosylmethionine (SAM) metabolic processes (10 DEGs and 3 DAPs), apoplastic ROS production (16 DEGs and 2 DAPs), and cell death (14 DEGs and 6 DAPs). Among the identified DEGs and DAPs, 63 significantly changed at both the transcript and protein levels during the experimental periods. In addition, the majority of these identified genes and proteins were expressed the most at the prime stage of stone cell differentiation, but their levels gradually decreased at the later stages.

Full-length transcriptome and targeted metabolome analyses provide insights into defense mechanisms of Malus sieversii against Agrilus mali.

Malus sieversii is the wild progenitor for many cultivars of domesticated apple and an important germplasm resource for breeding. However, this valuable species faces a significant threat in the areas north of the Tianshan Mountains in China, by the invasion of Agrilus mali, a destructive pest of apple trees belonging to the family Buprestidae. Our preliminary study has has shown that there may be resistance to this insect in M. sieversii plants in the field, but the corresponding molecular mechanisms remain unclear. In this study, we compared the response of insect-resistant and insect-susceptible plants of M. sieversii to insect feeding using full-length transcriptome and targeted metabolome. 112,103 non-chimeric full-length reads (FLNC) totaling 10.52 Gb of data were generating with Pacific Biosciences SingleMolecule, Real-Time (PacBio SMRT) sequencing. A total of 130.06 Gb data of long reads were acquired with an Illumina HiSeq. Function annotation indicated that the different expressed genes (DEGs) were mainly involved in signal transduction pathway of plant hormones and in the synthesis of compounds such as terpenes, quinones, flavonoids, and jasmonic acid. Through targeted metabolome analysis resistant strains showed higher levels of trans-cinnamic acid, caffeine and ferulic acid after pest infestation. This study helps to decipher the transcriptional changes and related signaling paths in M. sieversii after an insect feeding, which lays a foundation for further research on molecular mechanisms of insect resistance in apples.