Guanghua He

CHIMERIC FLORAL ORGANS1, Encoding a Monocot-Specific MADS Box Protein, Regulates Floral Organ Identity in Rice

The control of floral organ identity by homeotic MADS box genes is well established in eudicots. However, grasses have highly specialized outer floral organs, and the identities of the genes that regulate the highly specialized outer floral organs of grasses remain unclear. In this study, we characterized a MIKC-type MADS box gene, CHIMERIC FLORAL ORGANS (CFO1), which plays a key role in the regulation of floral organ identity in rice (Oryza sativa). The cfo1 mutant displayed defective marginal regions of the palea, chimeric floral organs, and ectopic floral organs. Map-based cloning demonstrated that CFO1 encoded the OsMADS32 protein. Phylogenetic analysis revealed that CFO1/OsMADS32 belonged to a monocot-specific clade in the MIKC-type MADS box gene family. The expression domains of CFO1 were mainly restricted to the marginal region of the palea and inner floral organs. The floral organ identity gene DROOPING LEAF (DL) was expressed ectopically in all defective organs of cfo1 flowers. Double mutant analysis revealed that loss of DL function mitigated some of the defects of floral organs in cfo1 flowers. We propose that the CFO1 gene plays a pivotal role in maintaining floral organ identity through negative regulation of DL expression.

MULTI-FLORET SPIKELET1 , Which Encodes an AP2/ERF Protein, Determines Spikelet Meristem Fate and Sterile Lemma Identity in Rice

MULTI-FLORET SPIKELET1 determines spikelet meristem fate and sterile lemma identity in rice. The spikelet is a unique inflorescence structure of grass. The molecular mechanism that controls the development of the spikelet remains unclear. In this study, we identified a rice (Oryza sativa) spikelet mutant, multi-floret spikelet1 (mfs1), that showed delayed transformation of spikelet meristems to floral meristems, which resulted in an extra hull-like organ and an elongated rachilla. In addition, the sterile lemma was homeotically converted to the rudimentary glume and the body of the palea was degenerated in mfs1. These results suggest that the MULTI-FLORET SPIKELET1 (MFS1) gene plays an important role in the regulation of spikelet meristem determinacy and floral organ identity. MFS1 belongs to an unknown function clade in the APETALA2/ethylene-responsive factor (AP2/ERF) family. The MFS1-green fluorescent protein fusion protein is localized in the nucleus. MFS1 messenger RNA is expressed in various tissues, especially in the spikelet and floral meristems. Furthermore, our findings suggest that MFS1 positively regulates the expression of LONG STERILE LEMMA and the INDETERMINATE SPIKELET1 (IDS1)-like genes SUPERNUMERARY BRACT and OsIDS1.

Transgenic indica rice expressing a bitter melon (Momordica charantia) class I chitinase gene (McCHIT1) confers enhanced resistance to Magnaporthe grisea and Rhizoctonia solani

McCHIT1 chitinase (DQ407723), a class I secretory endochitinase from bitter melon (Momordica charantia), had been demonstrated to enhance resistance against Phytophthora nicotianae and Verticillium wilt in transgenic tobacco and cotton. In order to obtain disease-resistant transgenic rice, McCHIT1 was transformed into a restorer line JinHui35 (Oryza sativa subsp. indica) by using the herbicide-resistance gene Bar as the selection marker. Transgenic rice lines and their progenies overexpressing the McCHIT1 gene showed enhanced resistance to Magnaporthe grisea (rice blast) and Rhizoctonia solani (sheath blight), two major fungal pathogens of rice. McCHIT1-transgenic rice confirmed the inheritance of the transgene and disease resistance to the subsequent generation. The T2 transformants exhibited significantly increased tolerance to M. grisea, with a 30.0 to 85.7 reduction in disease index, and R. solani, with a 25.0 to 43.0 reduction in disease index, based on that of the control as 100. These results indicated that over-expression of the McCHIT1 gene could lead to partial disease reduction against these two important pathogens in transgenic rice.

Assessment of purity of rice CMS lines using cpDNA marker

AFLP technique was used to analyse the polymorphism between rice cytoplasmic male sterility (CMS) line Jin2A and its maintainer Jin2B. A stable differential band was discovered, and sequence analysis showed that Jin2A contained a more tandem repeat of 6 base pairs (AGAAAA) than Jin2B. Further studies confirmed that the diversity came from cpDNA and occurred at three kinds of abortive cytoplasmic genotypes. Accordingly, specific primers were designed and utilized to assess the purity of rice CMS lines during multiplication with pollen fertility and seed setting rates of bagged panicles as control. The result indicated that this cpDNA locus could be utilized to precisely distinguish maintainer plants from rice CMS lines. PCR analysis was consistent with that from Grow-out test in CMS line seed purity assessment during multiplication, despite it was helpless in distinguishing F1 hybrids from CMS lines due to similar cytoplasms. Because of fewer hybrid and more maintainer off-plants, this cpDNA locus was still appropriate for seed purity assessment of rice CMS line during multiplication. This is first report that a marker on cpDNA could be utilized to assess the genetic purity of rice CMS lines with three abortive cytoplasmic genotypes.

Gene mapping related to yellow green leaf in a mutant line in rice (Oryza sativa L.)

A mutant, which derived from the restorer line Jinhui10 treated with EMS, showed completely yellow green leaves, and it had low chlorophyll content and poor agronomic characteristics during the growing stage. The F1 plants from the cross between normal × the mutant showed normal green leaves, and the segregation ratio of normal to yellow green leaves was 3 : 1 in F2 population. It indicated that the trait was controlled by a single recessive nuclear gene, temporarily designated asygl3. The geneygl3 was mapped between RM468 and RM3684 with genetic distances 8.4 cM and 1.8 cM on chromosome 3. This result would be used as genetic information for fine mapping and map-based cloning ofygl3 gene.

Physiological character and molecular mapping of leaf-color mutant wyv1 in rice (Oryza sativa L.)

The seed of an excellent indica restorer line Jinhui10 (Oryza sativa L. ssp. indica) was treated by ethyl methanesulfonate (EMS); a leaf-color mutant displaying distinct phenotype throughout development grown in paddy field was identified from the progeny. The mutant leaf showed white-yellow at seedling stage and then turned to yellow-green at tillering stage, after that, virescent color appeared until to maturity. The mutant was thus temporarily designed as wyv1. The chlorophyll contents decreased significantly and the changing was consistent with the chlorotic level of wyv1 leaves. Chlorophyll fluorescence kinetic parameters measured at the seedling stage showed that co-efficiency of photochemical quenching (qP), actual photosystem II efficiency (ΦPS II), electron transport rate (ETR) and initial chlorophyll fluorescence level (Fo), net photosynthetic rate (Pn) and maximum photochemical efficiency (Fv / Fm) significantly decreased in severe chlorotic leaf of the mutant compared with that of wild type. However, no significant differences were observed for Pn and Fv/Fm between virescent leaf and normal green leaf. Genetic analysis suggested that the mutant phenotype was controlled by a single recessive nuclear gene which was finally mapped between SSR marker Y7 and Y6 on rice chromosome 3 based on F2 population of Xinong1A / wyv1. Genetic distances were 0.06 cM and 0.03 cM respectively, and the physical distance was 84 kb according to the sequence of indica rice 9311. The results must facilitate map-based cloning and functional analysis of WYV1 gene.

Identification and Gene Mapping of a Novel Mutant supernumerary lodicules (snl) in Rice

In order to gain a better understanding of rice flower development, a rice flower mutant supernumerary lodicules (snl), which was identified from ethyl methane sulfonate (EMS)-treated Jinhui10 (Oryza sativa L. ssp. indica) was used in the present study. In the snl mutant, the palea obtained lemma identity, additional glume-like organs formed, lodicules increased and elongated, stamens decreased, and a few aberrant carpels formed. These phenotypes suggest that SNL is involved in the entire rice flower development. SNL was mapped between two simple sequence repeat markers RM3512 and RM1342 on chromosome 2, an approximate 800 kb region, and it co-segregated with SSR215. We conclude that SNL is a novel gene involved in flower development in rice. The present study will be useful for further cloning of the SNL gene, which will contribute to the elucidation of rice flower development.

Developmental genetics analysis for plant height in indica hybrid rice across environments.

The developmental genetics of plant height was analyzed from two groups of three-line indica hybrid rice at two environmental conditions based on the NCII design, using the additive-dominant developmental genetics models and the statistic methods. The results showed that the rice genotypes and environmental conditions could both affect plant height, and the effects of environment on plant height decreased gradually with plant development. Additive and dominant effects both governed the performance of plant height at all developmental stages. However, the degrees of effect varied among the rice genotypes. Moreover, the interaction between environments and genotypes also affected plant height. The genetic effects differed at most developmental stages. Furthermore, the expression of additive effect was more active than that of dominant effect. Conditional interaction effects with environment also influenced plant height during genetic development, especially at the early stage. Mid-parent heterosis (H(subscript MP)) increased gradually with the developmental stage of plant height, and maximized at the latest stage, whereas the heterosis over the better parent (H(subscript BP)) showed small differences among the genotypes, and kept stable at the later stage, with positive numeric value. At most developmental stages, conditional H(subscript MP) was positively significant, while conditional H(subscript BP) was negatively significant. All above results suggest that H(subscript MP) and H(subscript BP) have some new expressions in all developmental periods and the levels and directions are quite different.