The boron transporter SiBOR1 functions in cell wall integrity, cellular homeostasis, and panicle development in foxtail millet

Boron(B) is an essential micronutrient for vascular plant growth. Both B deficiency and toxicity can impair tissue development in diverse plant species, but little is known about the effect of B on reproductive panicle development and grain yield. In this study, a mutant of Setaria italica exhibiting necrotic panicle apices was identified and designated as sibor1. Sequencing revealed a candidate gene, Si BOR1, with a G-to-A alteration at the seventh exon. Knockout transgenic lines generated by clustered regularly interspaced short palindromic repeats and their associated protein-9 also had necrotic panicles, verifying the function of Si BOR1. Si BOR1 encoded a membrane-localized B efflux transporter, co-orthologous to the rice BOR1 protein. Si BOR1 was dominantly expressed in panicles and displayed a distinct expression pattern from those of its orthologs in other species. The induced mutation in Si BOR1 caused a reduction in the B content of panicle primary branches, and B deficiency-associated phenotypes such as thicker cell walls and higher cell porosity compared with Yugu 1. Transcriptome analysis indicated that differentially expressed genes involved in cell wall biogenesis, jasmonic acid synthesis, and programmed cell death response pathways were enriched in sibor1. q PCR analysis identified several key genes, including phenylalanine ammonia-lyase(Si PAL) and jasmonate-ZIM-domain(Si JAZ) genes, responsive to B-deficient conditions. These results indicate that Si BOR1 helps to regulate panicle primary branch development to maintain grain yield in S. italica. Our findings shed light on molecular mechanisms underlying the relationship between B transport and plant development in S. italica.

Comparison of Tissue Transglutaminase Activity During Young Panicle Development in Honglian-type Cytoplasmic Male Sterile Rice

To investigate the changes of tissue transglutaminase activity, the leaves and young panicles of rice at different developmental stages were excised from the Honglian-type cytoplasmic male sterile line, Yuetai A and its maintainer line, Yuetai B, respectively. An ELISA measurement protocol for tissue transglutaminase activity detection in rice was well established. The results indicated that the tissue transglutaminase activity was regulated positively by calcium cation, and the tissue transglutaminase activity in senescent leaves was remarkably higher than that in young leaves. No distinct difference was noted between Yuetai A and Yuetai B. Moreover, from the tetrad to binucleate stages the tissue transglutaminase activity increased gradually with the progression of the young panicle development and up to maximum at binucleate stage in Yuetai A. However, no similar changes were observed in Yuetai B. This indicates that the tissue transglutaminase is involved in cell programmed death in abortive pollen.

Decrement of Sugar Consumption in Rice Young Panicle Under High Temperature Aggravates Spikelet Number Reduction

Two rice genotypes Huanghuazhan(HHZ, heat-resistant) and IR36(heat-susceptible) were subjected to high-temperature(HT, 40℃) and normal-temperature(NT, 32℃) treatments at the spikelet differentiation stage. HT treatment inhibited spikelet differentiation, aggravated spikelet degeneration, reduced spikelet size, and disordered carbohydrate allocation. Meanwhile, HT treatment increased nonstructural carbohydrate content in leaves, but decreased that in stems and young panicles, and the same tendencies of sucrose and starch contents were observed in leaves and stem. However, HT treatment significantly increased the sucrose content and sharply decreased the glucose and fructose contents in young panicles. Lower activity levels of soluble acid invertase(EC3.2.1.26) and sucrose synthase(EC2.4.1.13) were observed under HT treatment. Moreover, HT treatment reduced the activities of key enzymes associated with glycolysis and the tricarboxylic acid cycle, which indicated sucrose consumption was inhibited in young panicles under HT treatment. Exogenous glucose and fructose applied under HT treatment increased the spikelet number more than exogenous sucrose. In conclusion, the results demonstrated that the reduction of spikelet number under high temperature was more affected by the decrease in sugar consumption than the blocking of sucrose transport. The impairment of sucrose hydrolysis was the main reason for the inhibition of sugar utilization.

NECK LEAF 1, a GATA type transcription factor, modulates organogenesis by regulating the expression of multiple regulatory genes during reproductive development in rice

In the monocot rice species Oryza sativa L., one of the most striking morphological processes during reproductive development is the concurrence of panicle development with the sequential elongation of upper internodes （UPIs）. To elucidate the underlying molecular mechanisms, we cloned the rice gene NECK LEAF 1 （NL1）, which when mutated results in delays in flowering time, smaller panicles with overgrown bracts and abnormal UPI elongation patterns. The NL1 gene encodes a GATA-type transcription factor with a single zinc finger domain, and its transcripts are de- tected predominantly in the bract primordia, which normally degenerate in the wild-type plants. Overexpression of NL1 in transgenic plants often gives rise to severe growth retardation, less vegetative phytomers and smaller leaves, suggesting that NL1 plays an important role in organ differentiation. A novel mutant allele of PLASTOCHRON1 （PLAD, a gene known to play a key role in regulating leaf initiation, was identified in this study. Genetic analysis demonstrated an interaction between nil and plal, with NL1 acting upstream of PLA1. The expression level and spatial pattern of PLA1 were found to be altered in the nil mutant. Furthermore, the expression of two regulators of flowering, Hd3a and OsMADS1, was also affected in the nil mutant. On the basis of these findings, we propose that NL1 is an intrinsic factor that modulates and coordinates organogenesis through regulating the expression of PLA1 and other regulatory genes during reproductive development in rice.

Coordinated Expression of Cytosolic and Chloroplastic Glutamine Synthetase During Reproductive Stage and Its Impact in GS1 RNAi Transgenic Rice

To understand the reallocation of organic nitrogen from leaf to the flower head of rice, the role of glutamine synthetase （GS） was investigated by characterizing GS1 RNAi transgenic rice, which revealed a significant reduction in panicle number and number of seeds per panicle. We observed the expression of GS isotypes at transcriptional and protein levels in flag leaves, leaf sheaths and panicles at three different flower development stages. The mRNA expression of GS1;1 was clearly suppressed in flag leaves, especially at the flowering stage. GS1 protein was barely detectable in flag leaves until the flowering stage, while GS1 protein was compromised in the leaf sheath and panicle, with transient expression of GS2 protein at the flowering stage. The glutamine level in transgenic plants was significantly reduced in both flag leaves and panicles, but ammonium was highly accumulated. The level of other amino acids, including aspartate and asparagine, tended to be higher in RNAi transgenic plants than the wild type plants during the reproductive stage. In addition, accumulation of toxic ammonium in panicles with low glutamine level might have caused low seed-setting in the transgenic rice. These results indicated that nitrogen reallocation was critical for panicle development, and that multiple GS isotypes functioned cooperatively to complete the rice life cycle when leaf nitrogen was remobilized to the developing reproductive organs.

Putative zeatin O-glucosyltransferase OscZOG1 regulates root and shoot development and formation of agronomic traits in rice

As a ubiquitous reaction, glucosylation controls the bioactivity of cytokinins in plant growth and development. Here we show that genetic manipulation of zeatin-O- glucosylation regulates the formation of important agronomic traits in rice by manipulating the expression of OscZOG1 gene, encoding a putative zeatin O-glucosyltransferase. We found that OscZOG~ was preferentially expressed in shoot and root meristematic tissues and nascent organs. The growth of lateral roots was stimulated in the overexpression lines, but inhibited in RNA interference lines. In shoots, knockdown of OscZOG1 expression by RNA interference significantly im- proved tillering, panicle branching, grain number per panicle and seed size, which are important agronomic traits for grain yield. In contrast, constitutive expression of OscZOG1 leads to negative effects on the formation of the grain-yielding traits with a marked increase in the accumulation levels of cis-zeatin O-glucoside （cZOG） in the transgenic rice plants. In this study,our findings demonstrate the feasibility of improving the critical yield-determinant agronomic traits, including tiller number, panicle branches, total grain number per panicle and grain weight by downregulating the expression level of OscZOG1. Our results suggest that modulating the levels of cytokinin glucosylation can function as a fine-tuning switch in regulating the formation of agronomic traits in rice.

A Rice Panicle Mutant Created by Transformation with an Antisense cDNA Library

A rice （Oryza sativa L.） mutant displaying defects in panicle development was identified among transformants in a transgenic mutagenlzed experiment using an antlsense cDNA library prepared from young rice panicles. In the mutant, the average splkelet number was reduced to 59.8 compared with 104.3 in wild-type plants. In addition, the seed-setting rate of the mutant was low （39.3%） owing to abnormal female development. Genetic analysis of T1 and T2 progeny showed that the traits segregated In a 3 （mutant） ： 1 （wild type） ratio and the mutation was cosegregated with the transgene. Southern blot and thermal asymmetric interlaced polymerase chain reaction analyses showed that the mutant had a single T-DNA insertion on chromosome 5, where no gene was tagged. Sequencing analysis found that the transgenic antisense cDNA was derived from a gene encoding an F-box protein in chromosome 7 with unidentified function. This and another four homologous genes encoding putative F-box proteins form a gene cluster. These results indicate that the phenotyplc mutations were most likely due to the silencing effect of the expressed transgenic antisense construct on the member（s） of the F-box gene cluster.