Cannabis sativa: origin and history, glandular trichome development, and cannabinoid biosynthesis

Is Cannabis a boon or bane?Cannabis sativa has long been a versatile crop for fiber extraction(industrial hemp),traditional Chinese medicine(hemp seeds),and recreational drugs(marijuana).Cannabis faced global prohibition in the twentieth century because of the psychoactive properties of △^(9)-tetrahydrocannabinol;however,recently,the perspective has changed with the recognition of additional therapeutic values,particularly the pharmacological potential of cannabidiol.A comprehensive understanding of the underlying mechanism of cannabinoid biosynthesis is necessary to cultivate and promote globally the medicinal application of Cannabis resources.Here,we comprehensively review the historical usage of Cannabis,biosynthesis of trichome-specific cannabinoids,regulatory network of trichome development,and synthetic biology of cannabinoids.This review provides valuable insights into the efficient biosynthesis and green production of cannabinoids,and the development and utilization of novel Cannabis varieties.

Genome-wide identification and characterization of ACBP gene family in Populus reveal salinity alkali-responsive profiles

Acyl-CoA-binding proteins(ACBPs)are important for the transport of acyl groups for macro molecular biosynthesis involved in plant growth,development,and diverse stress(e.g.,cold,drought,salinity,and heavy metals)responses.Here,we report the phylogeny and characteristics of the ACBP family in the woody plant Populus trichocarpa.Eight genes encoding ACBP proteins were identified,and they are distributed on eight chromosomes in P.trichocarpa.These PtACBP genes were divided into four subgroups according to gene structure,conserved motifs and phylogenetic relationship.Promoter analysis revealed that cis-elements were related to stress response,phytohormone response,and physical and reproductive growth regulation.Expression levels of PtACBP genes varied among different organs,with the highest expression in leaves and the lowest in stems.Quantitative real-time PCR(qRT-PCR)analysis showed that under salinity-alkali stresses(i.e.,200 mM NaCl,75 mM Na2CO3,and 100 mM NaHCO3),four(PtACBP1,PtACBP3,PtACBP4 and PtACBP8)of eight PtACBP genes were significantly induced in roots and leaves.These data provide a comprehensive analysis of the ACBPs family in P.trichocarpa,which could be useful for gene function analyses.

Genome-Wide Analysis of the KANADI Gene Family and Its Expression Patterns under Different Nitrogen Concentrations Treatments in Populus trichocarpa

KANADI(KAN)is a plant-specific gene that controlled the polarity development of lateral organs.It mainly acted on the abaxial characteristics of plants to make the lateral organs asymmetrical.However,it had been less identified in woody plants.In this study,the members of the KAN gene family in Populus trichocarpa were identified and analyzed using the bioinformatics method.The results showed that a total of 8 KAN family members were screened out,and each member contained the unique GARP domain and conserved region of the family proteins.Phylogenetic analysis and their gene structures revealed that all KAN genes from P.trichocarpa,Arabidopsis thaliana,and Nicotiana benthamiana could be divided into four subgroups,while the eight genes in P.trichocarpa were classified into three subgroups,respectively.The analysis of tissue-specific expression indicated that PtKAN1 was highly expressed in young leaves,PtKAN6 was highly expressed in young leaves and mature leaves,PtKAN2,PtKAN5,and PtKAN7 were highly expressed in nodes and internodes,PtKAN8 was highly expressed in roots,and PtKAN3 and PtKAN4 showed low expression levels in all tissues.Among them,PtKAN2 and PtKAN6,and PtKAN4 and PtKAN5 might have functional redundancy.Under high nitrogen concentrations,PtKAN2 and PtKAN8 were highly expressed in mature stems and leaves,respectively,while PtKAN4,PtKAN5,and PtKAN7 were highly expressed in roots.This study laid a theoretical foundation for further study of the KAN genemediated nitrogen effect on root development.

Nitrogen application and intercropping change microbial community diversity and physicochemical characteristics in mulberry and alfalfa rhizosphere soil

Intercropping of mulberry(Morus alba L.)and alfalfa(Medicago sativa L.)is a new forestry-grass compound model in China,which can provide high forage yields with high protein.Nitrogen application is one of the important factors determining the production and quality of this system.To elucidate the advantages of intercropping and nitrogen application,we analyzed the changes of physicochemical properties,enzyme activities,and microbial communities in the rhizosphere soil.We used principal components analysis(PCA)and redundancy discriminators analysis to clarify the relationships among treatments and between treatments and environmental factors,respectively.The results showed that nitrogen application significantly increased pH value,available nitrogen content,soil water content(SWC),and urea(URE)activity in rhizosphere soil of monoculture mulberry.In contrast,intercropping and intercropping+N significantly decreased pH and SWC in mulberry treatments.Nitrogen,intercropping and intercropping+N sharply reduced soil organic matter content and SWC in alfalfa treatments.Nitrogen,intercropping,and intercropping+N increased the values of McIntosh diversity(U),Simpson diversity(D),and Shannon-Weaver diversity(H’)in mulberry treatments.However,PC A scatter plots showed clustering of monoculture mulberry with nitrogen(MNE)and intercropping mulberry without nitrogen(M0).Intercropping reduced both H’and D but nitrogen application showed no effect on diversity of microbial communities in alfalfa.There were obvious differences in using the six types of carbon sources between mulberry and alfalfa treatments.Nitrogen and intercropping increased the numbers of sole carbon substrate in mulberry treatments where the relative use rate exceeded 4%.While the numbers declined in alfalfa with nitrogen and intercropping.RDA indicated that URE was positive when intercropping mulberry was treated with nitrogen,but was negative in monoculture alfalfa treated with nitrogen.Soil pH and SWC were positive with mulberry treatments but were negative with alfalfa treatments.Intercropping with alfalfa benefited mulberry in the absence of nitrogen application.Intercropping with alfalfa and nitrogen application could improve the microbial community function and diversity in rhizosphere soil of mulberry.The microbial community in rhizosphere soil of mulberry and alfalfa is strategically complementary in terms of using carbon sources.

PuHox52 promotes coordinated uptake of nitrate,phosphate, and iron under nitrogen deficiency in Populus ussuriensis

It is of great importance to better understand how trees regulate nitrogen(N) uptake under N deficiency conditions which severely challenge afforestation practices, yet the underlying molecular mechanisms have not been well elucidated. Here,we functionally characterized PuHox52, a Populus ussuriensis HD-ZIP transcription factor, whose overexpression greatly enhanced nutrient uptake and plant growth under N deficiency. We first conducted an RNA sequencing experiment to obtain root transcriptome using PuHox52-overexpression lines of P. ussuriensis under low N treatment. We then performed multiple genetic and phenotypic analyses to identify key target genes of PuHox52 and validated how they acted against N deficiency under PuHox52 regulation.PuHox52 was specifically induced in roots by N deficiency, and overexpression of PuHox52promoted N uptake, plant growth, and root development. We demonstrated that several nitrate-responsive genes(PuNRT1.1, PuNRT2.4,PuCLC-b, PuNIA2, PuNIR1, and PuNLP1),phosphate-responsive genes(PuPHL1A and PuPHL1B), and an iron transporter gene(PuIRT1) were substantiated to be direct targets of PuHox52. Among them, PuNRT1.1, PuPHL1A/B, and PuIRT1 were upregulated to relatively higher levels during PuHox52-mediated responses against N deficiency in PuHox52-overexpression lines compared to WT. Our study revealed a novel regulatory mechanism underlying root adaption to N deficiency where PuHox52 modulated a coordinated uptake of nitrate, phosphate, and iron through 'PuHox52-PuNRT1.1', 'PuHox52-PuPHL1A/PuPHL1B', and'PuHox52-PuIRT1' regulatory relationships in poplar roots.

A high-quality genome sequence of alkaligrass provides insights into halophyte stress tolerance

Alkaligrass(Puccinellia tenuiflora) is a monocotyledonous halophytic forage grass widely distributed in Northern China. It belongs to the Gramineae family and shares a close phylogenetic relationship with the cereal crops, wheat and barley. Here, we present a high-quality chromosome-level genome sequence of alkaligrass assembled from Illumina, Pac Bio and 10× Genomics reads combined with genome-wide chromosome conformation capture(Hi-C) data. The ~1.50 Gb assembled alkaligrass genome encodes 38,387 protein-coding genes, and 54.9% of the assembly are transposable elements, with long terminal repeats being the most abundant. Comparative genomic analysis coupled with stress-treated transcriptome profiling uncovers a set of unique saline-and alkaline-responsive genes in alkaligrass. The high-quality genome assembly and the identified stress related genes in alkaligrass provide an important resource for evolutionary genomic studies in Gramineae and facilitate further understanding of molecular mechanisms underlying stress tolerance in monocotyledonous halophytes. The alkaligrass genome data is freely available at http://xhhuanglab.cn/data/alkaligrass.html.

OsMKKK70 regulates grain size and leaf angle in rice through the OsMKK4-OsMAPK6-OsWRKY53 signaling pathway

Grain size and leaf angle are key agronomic traits that determine final yields in rice.However,the underlying molecular mechanisms are not well understood.Here we demonstrate that the Oryza sativa Mitogen Activated Protein Kinase Kinase Kinase OsMKKK70 regulates grain size and leaf angle in rice.Overexpressing OsMKKK70 caused plants to produce longer seeds.The osmkkk62/70 double mutant and the osmkkk55/62/70 triple mutant displayed significantly smaller seeds and a more erect leaf angle compared to the wild type,indicating that OsMKKK70 functions redundantly with its homologs Os MKKK62 and Os MKKK55.Biochemical analysis demonstrated that OsMKKK70 is an active kinase and that OsMKKK70 interacts with Os MKK4 and promotes Os MAPK6 phosphorylation.In addition,the osmkkk62/70 double mutant showed reduced sensitivity to Brassinosteroids(BRs).Finally,overexpressing constitutively active Os MKK4,Os MAPK6,and Os WRKY53 can partially complement the smaller seed size,erect leaf,and BR hyposensitivity of the osmkkk62/70 double mutant.Taken together,these findings suggest that OsMKKK70 might regulate grain size and leaf angle in rice by activating Os MAPK6 and that OsMKKK70,Os MKK4,Os MAPK6,and Os WRKY53 function in a common signaling pathway that controls grain shape and leaf angle.

bZIP71 delays flowering by suppressing Ehd1 expression in rice

Flowering time is a fundamental factor determining the global distribution and final yield of rice(Oryza sativa).Although diverse flowering time genes have been reported in this crop,the transcriptional regulation of its key flowering genes are poorly understood.Here,we report that a basic leucine zipper transcription factor,bZIP71,functions as a flowering repressor.The overexpression of bZIP71 delays flowering,while the bzip71 mutant flowers early in both long-day and short-day conditions.A genetic analysis showed that the regulation of flowering by bZIP71 might be independent of Heading date 2(Hd2),Hd4,and Hd5.Importantly,bZIP71 directly associates with the Early heading date 1(Ehd1)promoter and represses its transcription,and genetically the function of bZIP71 is impaired in the ehd1 mutant.Moreover,bZIP71 interacts with major components of polycomb repressive complex 2(PRC2),SET domain group protein 711(SDG711),and Fertilization independent endosperm 2(FIE2),through which bZIP71 regulates the H3K27me3 level of Ehd1.Taken together,we present a transcriptional regulatory mechanism in which bZIP71 enhances the H3K27me3 level of Ehd1 and transcriptionally represses its expression,which not only offers a novel insight into a flowering pathway,but also provides a valuable putative target for the genetic engineering and breeding of elite rice cultivars.