The OsMYC2-JA feedback loop regulates diurnal flower-opening time via cell wall loosening in rice.

Diurnal flower-opening time (DFOT), the time of spikelet opening during the day, is an important trait for hybrid rice (Oryza sativa L.) seed production. Hybrids between indica and japonica rice varieties have strong heterosis, but the parental lines usually have different, nonoverlapping DFOTs. This reduces the success of hybrid seed production in crosses between indica and japonica subspecies, thus hindering the utilization of indica and japonica inter-subspecies heterosis. However, little is known about the molecular mechanisms regulating DFOT in rice. Here, we obtained japonica rice lines with a DFOT 1.5 h earlier than the wild type by overexpressing OsMYC2, a gene encoding a key transcription factor in the jasmonate (JA) signaling pathway. OsMYC2 is activated by JA signaling and directly regulates the transcription of genes related to JA biosynthesis and cell wall metabolism. Overexpressing OsMYC2 led to significantly increased JA contents and decreased cellulose and hemicellulose contents in lodicule cells, as well as the softening of lodicule cell walls. This may facilitate the swelling of lodicules, resulting in early diurnal flower-opening. These results suggest that the OsMYC2-JA feedback loop regulates DFOT in rice via cell wall remodeling. These findings shed light on the understanding of regulatory mechanism of the DFOT of plants, which should promote the development of indica and japonica varieties suitable for hybrid rice breeding.

Is the time of anthesis in rice (Oryza sativa) influenced by photoperiod?

Photoperiod sensitivity in rice cultivars is defined when the cultivar begins anthesis on a relatively invariant date, varying by < 7 days, regardless of the date of sowing or germination. While the date of flowering in photoperiod sensitive (PPS) rice cultivars is characteristically determined by the day length, especially during the short-day season (September-December), the response of the flower opening time (FOT) to photoperiod remains hitherto unexplored. This paper examines whether day length restrains year-to-year variation in FOT in PPS cultivars. We examined 105 PPS and 173 photoperiod insensitive (PPI) cultivars grown in different years and estimated their year-to-year FOT difference (or FOTD) and the year-to-year difference of sunrise to anthesis duration (or SADD). Wilcoxon signed rank test and bootstrap test were then performed to test whether these descriptors significantly differed between PPS and PPI groups of cultivars. The means of FOTD and SADD were detected to be significantly less in the PPS group than in the PPI group of cultivars, indicating significantly lesser variability of FOT in PPS than in PPI cultivars. This is the first report of a strong restraining influence of photoperiod on FOT variability in PPS cultivars.

Controlling diurnal flower-opening time by manipulating the jasmonate pathway accelerates development of indica-japonica hybrid rice breeding.

Inter-subspecific indica-japonica hybrid rice (Oryza sativa) has the potential for increased yields over traditional indica intra-subspecies hybrid rice, but limited yield of F1 hybrid seed production (FHSP) hinders the development of indica-japonica hybrid rice breeding. Diurnal flower-opening time (DFOT) divergence between indica and japonica rice has been a major contributing factor to this issue, but few DFOT genes have been cloned. Here, we found that manipulating the expression of jasmonate (JA) pathway genes can effectively modulate DFOT to improve the yield of FHSP in rice. Treating japonica cultivar Zhonghua 11 (ZH11) with methyl jasmonate (MeJA) substantially advanced DFOT. Furthermore, overexpressing the JA biosynthesis gene OPDA REDUCTASE 7 (OsOPR7) and knocking out the JA inactivation gene CHILLING TOLERANCE 1 (OsHAN1) in ZH11 advanced DFOT by 1- and 2-h respectively; and knockout of the JA signal suppressor genes JASMONATE ZIM-DOMAIN PROTEIN 7 (OsJAZ7) and OsJAZ9 resulted in 50-min and 1.5-h earlier DFOT respectively. The yields of FHSP using japonica male-sterile lines GAZS with manipulated JA pathway genes were significantly higher than that of GAZS wildtype. Transcriptome analysis, cytological observations, measurements of elastic modulus and determination of cell wall components indicated that the JA pathway could affect the loosening of the lodicule cell walls by regulating their composition through controlling sugar metabolism, which in turn influences DFOT. This research has vital implications for breeding japonica rice cultivars with early DFOT to facilitate indica-japonica hybrid rice breeding.

Morning and evening alarm of the circadian clock for flower opening times in Hemerocallis.

Previous studies demonstrated that flower opening time (FOT) is a stable trait and precisely controlled by a circadian clock responsive to the environment. It plays a vital role in improving fertility. Hemerocallis spp. has different FOTs divided into two types: nocturnal and diurnal. To explore the regulatory mechanisms of their FOTs, we carried out a transcriptome sequencing experiment at different developmental stages of an F1 population with different FOTs. 55,883 unigenes were obtained, and 9234 differential genes were identified. Co-expression was analyzed by K-means clustering and weighted gene co-expression network analysis. Results showed that after entering reproductive growth, two FOT types of Hemerocallis had increased expression of genes related to photosynthetic metabolism and sensitivity to environmental response such as light and hormone signal transmission. Circadian rhythm-related activities were enriched in hub genes during the flowering stage. Genes showing differential expression between the two Hemerocallis groups were related to environmental response and photosynthesis pathways. Putative circadian clock genes displayed differences in expression across the flower opening stage in both groups of Hemerocallis. Twenty-three key circadian clock genes were identified, which related to sensitivity to light signal input and gating. These genes might closely relate to FOTs in Hemerocallis.

Effect of the light and dark conditions on flower opening time between cultivated rice (Oryza sativa) and a near-isogenic early-morning flowering line.

Flower opening time (FOT) is affected by genetic and environmental factors, but little is known about the effect of light and dark conditions on FOT in cereal crops. FOT of an indica rice cultivar, IR64, and its near-isogenic line carrying a QTL for an early-morning flowering trait (IR64+qEMF3) were investigated in a natural-light and temperature-controlled small greenhouse by exposing either the panicle or stem or both plant organs to different light and dark conditions. FOT did not change in either genotype when panicles were exposed to light. A large difference in FOT was found between genotypes when panicles were exposed to dark conditions; no flower opening was observed in IR64, whereas flower opening was delayed but observed in IR64+qEMF3. These results suggest that the panicle is the organ that perceives light for flower opening in both genotypes, whereas the light requirement to reach flower opening was quite different between genotypes. Flower opening of IR64 occurred concomitantly with elongation of anther filament in the light after the dark treatment stopped, whereas approximately half of flowering of IR64+qEMF3 resulted in apparent cleistogamy even during dark treatment. An extended duration of the dark treatment until 1730H (30-50 min before sunset) made FOT of IR64 spikelets on the next day shifted to a time as early as that of IR64+qEMF3, with significant advancement of FOT compared to the control IR64 spikelets. Our results indicated that different flowering responses to light and dark conditions exist between IR64 and IR64+qEMF3. These findings provide clues for understanding the unique genetic controls of flowering in an EMF line in rice. This study also showed evidence that artificial light environments can shift the FOT of IR64 to that of IR64+qEMF3.

Neural Net Classification Combined With Movement Analysis to Evaluate Setaria viridis as a Model System for Time of Day of Anther Appearance.

In many plant species, the time of day at which flowers open to permit pollination is tightly regulated. Proper time of flower opening, or Time of Day of Anther Appearance (TAA), may coordinate flowering opening with pollinator activity or may shift temperature sensitive developmental processes to cooler times of the day. The genetic mechanisms that regulate the timing of this process in cereal crops are unknown. To address this knowledge gap, it is necessary to establish a monocot model system that exhibits variation in TAA. Here, we examine the suitability of Setaria viridis, the model for C4 photosynthesis, for such a role. We developed an imaging system to monitor the temporal regulation of growth, flower opening time, and other physiological characteristics in Setaria. This system enabled us to compare Setaria varieties Ames 32254, Ames 32276, and PI 669942 variation in growth and daily flower opening time. We observed that TAA occurs primarily at night in these three Setaria accessions. However, significant variation between the accessions was observed for both the ratio of flowers that open in the day vs. night and the specific time of day where the rate is maximal. Characterizing this physiological variation is a requisite step toward uncovering the molecular mechanisms regulating TAA. Leveraging the regulation of TAA could provide researchers with a genetic tool to improve crop productivity in new environments.

qEMF3, a novel QTL for the early-morning flowering trait from wild rice, Oryza officinalis, to mitigate heat stress damage at flowering in rice, O. sativa.

A decline in rice (Oryza sativa L.) production caused by heat stress is one of the biggest concerns resulting from future climate change. Rice spikelets are most susceptible to heat stress at flowering. The early-morning flowering (EMF) trait mitigates heat-induced spikelet sterility at the flowering stage by escaping heat stress during the daytime. We attempted to develop near-isogenic lines (NILs) for EMF in the indica-type genetic background by exploiting the EMF locus from wild rice, O. officinalis (CC genome). A stable quantitative trait locus (QTL) for flower opening time (FOT) was detected on chromosome 3. A QTL was designated as qEMF3 and it shifted FOT by 1.5-2.0 h earlier for cv. Nanjing 11 in temperate Japan and cv. IR64 in the Philippine tropics. NILs for EMF mitigated heat-induced spikelet sterility under elevated temperature conditions completing flower opening before reaching 35°C, a general threshold value leading to spikelet sterility. Quantification of FOT of cultivars popular in the tropics and subtropics did not reveal the EMF trait in any of the cultivars tested, suggesting that qEMF3 has the potential to advance FOT of currently popular cultivars to escape heat stress at flowering under future hotter climates. This is the first report to examine rice with the EMF trait through marker-assisted breeding using wild rice as a genetic resource.