Summary: Red rice contains high levels of proanthocyanidins and anthocyanins, which have been recognized as health‐promoting nutrients. The red coloration of rice grains is controlled by two complementary genes, Rc and Rd. The RcRd genotype produces red pericarp in wild species Oryza rufipogon, whereas most cultivated rice varieties produce white grains resulted from a 14‐bp frame‐shift deletion in the seventh exon of the Rc gene. In the present study, we developed a CRISPR/Cas9‐mediated method to functionally restore the recessive rc allele through reverting the 14‐bp frame‐shift deletion to in‐frame mutations in which the deletions were in multiples of three bases, and successfully converted three elite white pericarp rice varieties into red ones. Rice seeds from T₁ in‐frame Rc lines were measured for proanthocyanidins and anthocyanidins, and high accumulation levels of proanthocyanidins and anthocyanidins were observed in red grains from the mutants. Moreover, there was no significant difference between wild‐type and in‐frame Rc mutants in major agronomic traits, indicating that restoration of Rc function had no negative effect on important agronomic traits in rice. Given that most white pericarp rice varieties are resulted from the 14‐bp deletion in Rc, it is conceivable that our method could be applied to most white pericarp rice varieties and would greatly accelerate the breeding of new red rice varieties with elite agronomic traits. In addition, our study demonstrates an effective approach to restore recessive frame‐shift alleles for crop improvement.

Background: Homegardens are ecologically and culturally important systems for cultivating medicinal plants for wellbeing by healers and farmers in Naxi communities of the Sino Himalayan region. The cultivation of medicinal plants in Naxi communities and associated ethnomedical knowledge base for maintaining and utilizing these resources is at risk with expanded commercialization of natural resources, development policies and rapid socio-economic change in China. Research is needed to understand the medicinal plant species maintained in Naxi homegardens, their use and contribution to community wellbeing, and how these practices and knowledge base varies between Naxi healers and farmers in order to develop plans for biodiversity conservation and preservation of ethnomedical practices. The main objective of this study is to document and compare medicinal plant species in Naxi homegardens and associated ethnomedical knowledge between Naxi healers and farmers.Methods: Ethnobotanical homegarden surveys were conducted with three Naxi healers and 28 farmer households in two Naxi communities in Lijiang Prefecture in Northwest Yunnan Province of China. Surveys included inventories of medicinal plants in homegardens and semi-structured interviews with homegarden managers to document traditional medicinal uses of inventoried plants. Inventoried plants were classified into 13 ‘usage categories’ of medical condition groupings that impact a system of the body. Finally, plant species richness was calculated for each homegarden and species richness was compared between healers and farmers as well as between study sites using a Least Square Means Tukey HSD function.Results: Ethnobotanical surveys at the study sites found that 13% of households rely exclusively on traditional Naxi medicine, 26% exclusively use Western medicine and 61% use a combination of traditional Naxi and Western medicine. A total of 106 medicinal plants were inventoried in Naxi homegardens representing 50 botanical families. Over 85% of inventoried medicinal plants were herbaceous. The most represented families were Asteraceae (12.8%), Ranunculaceae (8.3%), Apiaceae (8.3%), and Polygonaceae (7.3%). The primary medical functions of inventoried plants were to treat inflammation (73 species), circulatory system disorders (62), nervous system disorders (41), detoxification (39), digestive system disorders (33), muscular-skeletal system disorders (26), genitourinary system disorders (26), skin conditions (23), respiratory systems disorders (22), and cold and flu (20). Local herbal experts maintained greater medicinal plant species richness in their homegardens compared to local farmers as well as had greater knowledge of medicinal functions of plants. Healers maintained medicinal plants primarily for healing while farmer households maintained approximately 90% of the medicinal plants in their homegardens for commercialization and the remaining for household healthcare.Conclusions: This study highlights the importance of biodiversity and traditional ecological and medical knowledge for human wellbeing and livelihoods in Naxi communities. Conservation efforts and policies are necessary to preserve the ecological and cultural base that maintains medicinal plant use by both healers and farmers in Naxi homegardens of the Sino Himalayan region.

The transcription factor Woolly (Wo) and its downstream gene CycB2 have been shown to regulate trichome development in tomato (Solanum lycopersicum). It has been demonstrated that only the gain-of-function allele of Slwo (SlWo^V, the Slwo woolly motif mutant allele) can increase the trichome density; however, it remains unclear why the two alleles function differently in trichome development. In this study, we used Nicotiana benthamiana as a model and cloned the homologues of Slwo and SlCycB2 (named Nbwo and NbCycB2). We also constructed a Nbwo gain-of-function allele with the same mutation site as SlWo^V (named NbWo^V). We found that both Nbwo and NbWo^V directly regulate NbCycB2 and their own expression by binding to the promoter of NbCycB2 and their own genomic sequences. As form of a feedback regulation, NbCycB2 negatively regulates trichome formation by repressing Nbwo activity at the protein level. We also found that mutations in the Nbwo woolly motif can prevent repression of NbWo^V by NbCycB2, which results in a significant increase in the amount of active Nbwo proteins and in increases in trichome density and the number of branches. Our results reveal a novel reciprocal regulation mechanism between NbCycB2 and Nbwo during trichome formation in N. benthamiana.