Citation: Wang W, Zhang W, Jamil M, Tu J and Huang L (2024) Editorial: Molecular and genetic mechanisms of plant architecture regulation. Front. Plant Sci. 15:1421197. doi: 10.3389/fpls.2024.1421197

To allay excessive public concern about the safety of transgenic foods, and to optimize insect-resistant genes expression to delay the evolution of resistance in pests, we developed a promising strategy to fuse the GOI (gene of interest) with OsrbcS (rice small subunit of ribulose bisphosphate carboxylase/oxygenase) in transgenic rice, which acted as a carrier, driven by the OsrbcS native promoter to sequester its expression in green tissues. Using eYFP as a trial, we reported a high-level accumulation of eYFP in green tissue and almost none in the seed and root of the fused construct compared to the non-fused construct. After applying this fusion strategy in insect-resistant rice breeding, recombinant OsrbcS-Cry1Ab/Cry1Ac expressed rice plants conferred high resistance to leaffolders and striped stem borers, among which two single-copy lines possessed normal agronomic performance in the field. Specifically, Cry1Ab/Cry1Ac protein levels in single-copy construct transgenic lines ranged from 1.8 to 11.5 µg g−1 in the leaf, higher than the Actin I promoter-driven control, T51-1, about 1.78 µg g−1 in the leaf, but negligible (only 0.00012–0.00117 µg g−1) in endosperm by ELISA analysis. Our study provided a novel approach to creating Cry1Ab/Cry1Ac-free endosperm rice with a high level of insect-resistant protein in green tissues through the simultaneous usage of the OsrbcS promoter and OsrbcS as a fusion partner.

This research focused on cadmium (Cd), which negatively affects plant growth and auxin hemostasis. In plants, many processes are indirectly controlled through the expression of certain genes due to the secretion of bacterial auxin, as indole-3-acetic acid (IAA) acts as a reciprocal signaling molecule in plant-microbe interaction. The aim of current studies was to investigate responsible genes in rice for plant-microbe interaction and lateral root development due to the involvement of several metabolic pathways. Studies revealed that GH3-2 interacts with endogenous IAA in a homeostasis manner without directly providing IAA. In rice, indole-3-pyruvate decarboxylase (IPDC) transgenic lines showed a 40% increase in lateral roots. Auxin levels and YUCCA (auxin biosynthesis gene) expression were monitored in osaux1 mutant lines inoculated with Bacillus cereus exposed to Cd. The results showed an increase in root hairs (RHs) and lateral root density, changes in auxin levels, and expression of the YUCCA gene. B. cereus normalizes the oxidative stress caused by Cd due to the accumulation of O 2 - and H2O2 in osaux1 mutant lines. Furthermore, the inoculation of B. cereus increases DR5:GUS expression, indicating that bacterial species have a positive role in auxin regulation. Thus, the current study suggests that B. cereus and IPDC transgenic lines increase the RH development in rice by interacting with IAA synthetase genes in the host plant, alleviating Cd toxicity and enhancing plant defense mechanisms.

In models of competition in which space is treated as a continuum, and population size as continuous, there are no limits to the number of species that can coexist. For a finite number of sites, N, the results are different. The answer will, of course, depend on the model used to ask the question. In the Tilman-May-Nowak ordinary differential equation model, the number of species is asymptotically C log N with most species packed in at the upper end of the competitive hierarchy. In contrast, for metapopulation models with discrete individuals and stochastic spatial systems with various competition neighborhoods, we find a traditional species area relationship CN(a), with no species clumping along the phenotypic gradient. The exponent a is larger by a factor of 2 for spatially explicit models. In words, a spatial distribution of competitors allows for greater diversity than a metapopulation model due to the effects of recruitment limitation in their competition.

In order to create novel germplasm resources for breeding heat tolerant variety, we transferred a dominant allele OsHTAS, previously characterized and cloned from a high-temperature tolerant local variety HT54, which was collected from the rice production area of southern China, into a high- temperature sensitive intermediate breeding line HT13 through six rounds of successive backcross by using marker-assisted selection. The molecular analysis showed that the recovery of genetic background of a resultant near isogenic line (NIL), MHT13, was around 99.8%. The OsHTAS gene introduced in the MHT13 expressed normally in the HT13 genetic background, mediating heat tolerance and phenotype similar to those of the donor parent HT54. The major agronomic traits of MHT13 resembled those of the recurrent parent HT13. Moreover, MHT13 had high general combining ability and its rice quality reached the grade 3 standard of edible high-quality rice issued by Ministry of Agriculture of the People's Republic of China, which greatly improved its application value in rice production.

Chloroplasts are crucial organelles for the generation of fatty acids and starch required for plant development. Nascent polypeptide-associated complex (NAC) proteins have been implicated in development as transcription factors. However, their chaperone roles in chloroplasts and their relationship with pollen development in plants remain to be elucidated. Here, we demonstrated that Osj10gBTF3, a NAC protein, regulates pollen and chloroplast development in rice by coordinating with a Hsp90 family chaperone OsHSP82 to mediate chloroplast import. Knockout of Osj10gBTF3 affects pollen and chloroplast development and significantly reduces the accumulation of fertility-related chloroplast protein OsPPR676. Both Osj10gBTF3 and OsHSP82 interact with OsPPR676. Interestingly, the interaction between OsHSP82 and OsPPR676 is only found in the cytoplasm, while the interaction between Osj10gBTF3 and OsPPR676 also occurs inside the chloroplast. The chloroplast stroma chaperone OsCpn60 can also be co-precipitated with Osj10gBTF3, but not with OsHSP82. Further investigation indicates that Osj10gBTF3 enters the chloroplast stroma possibly through the inner chloroplast membrane channel protein Tic110 and then recruits OsCpn60 for the folding or assembly of OsPPR676. Our results reveal a chaperone role of Osj10gBTF3 in chloroplast import different from Hsp90 and provide a link between chloroplast transport and pollen development in rice.

The initiation stage of protein biosynthesis is a sophisticated process tightly regulated by numerous initiation factors and their associated components. However, the mechanism underlying translation initiation has not been completely understood in rice. Here, we showed knock-out mutation of the rice eukaryotic translation initiation factor 3 subunit h (OseIF3h) resulted in plant growth retardation and seed-setting rate reduction as compared to the wild type. Further investigation demonstrated an interaction between OseIF3h and OsMTA2 (mRNA adenosine methylase 2), a rice homolog of METTL3 (methyltransferase-like 3) in mammals, which provided new insight into how N6-methyladenosine (m6A) modification of messenger RNA (mRNA) is engaged in the translation initiation process in monocot species. Moreover, the RIP-seq (RNA immunoprecipitation sequencing) data suggested that OseIF3h was involved in multiple biological processes, including photosynthesis, cellular metabolic process, precursor metabolites, and energy generation. Therefore, we infer that OseIF3h interacts with OsMTA2 to target a particular subset of genes at translational level, regulating plant growth and pollen development.

P-subfamily PPR protein OsPPR939, which can be phosphorylated by OsS6K1, regulates plant growth and pollen development by involving in the splicing of mitochondrial nad5 introns 1, 2, and 3. In land plants, pentatricopeptide repeat (PPR) proteins play key roles in mitochondrial group II intron splicing, but how these nucleus-encoded proteins are imported into mitochondria is unknown. To date, a few PPR proteins have been characterized in rice (Oryza sativa). Here, we demonstrate that the mitochondrion-localized P-subfamily PPR protein OsPPR939 is required for the splicing of nad5 introns 1, 2, and 3 in rice. Complete knockout or partial disruption of OsPPR939 function resulted in different degrees of growth retardation and pollen sterility. The dramatically reduced splicing efficiency of these introns in osppr939-4 and osppr939-5 led to reduced mitochondrial complex I abundance and activity and enhanced expression of alternative respiratory pathway genes. Complementation with OsPPR939 rescued the defective plant morphology of osppr939-4 and restored its decreased splicing efficiency of nad5 introns 1, 2, and 3. Therefore, OsPPR939 plays crucial roles in plant growth and pollen development by splicing mitochondrial nad5 introns 1, 2, and 3. More importantly, the 12th amino acid Ser in the N-terminal targeting sequence of OsPPR939 is phosphorylated by OsS6K1, and truncated OsPPR939 with a non-phosphorylatable S12A mutation in its presequence could not be imported into mitochondria, suggesting that phosphorylation of this amino acid plays an important role in the mitochondrial import of OsPPR939. To our knowledge, the 12th residue Ser on OsPPR939 is the first experimentally proven phosphorylation site in PPR proteins. Our results provide a basis for investigating the regulatory mechanism of PPR proteins at the post-translational level.

Cytidine to uridine (C-to-U) RNA editing is an important type of substitutional RNA modification and is almost omnipresent in plant chloroplasts and mitochondria. In rice mitochondria, 491 C-to-U editing sites have been identified previously, and case studies have elucidated the function of several C-to-U editing sites in rice, but the functional consequence of most C-to-U alterations needs to be investigated further. Here, by means of Sanger sequencing and publicly available RNA-seq data, we identified a total of 569 C-to-U editing sites in rice mitochondria-encoded open reading frames (ORFs), 85.41% of these editing sites were observed on the first or the second base of a codon, resulting in the alteration of encoded amino acid. Moreover, we found some novel editing sites and several inaccurately annotated sites which may be functionally important, based on the highly conserved amino acids encoded by these edited codons. Finally, we annotated all 569 C-to-U RNA editing sites in their biological context. More precise information about C-to-U editing sites in rice mitochondria-encoded ORFs will facilitate our investigation on the function of C-to-U editing events in rice and also provide a valid benchmark from rice for the analysis of mitochondria C-to-U editing in other plant species.

EMP5 and OTP72 are PPR proteins involved in mitochondrial same site RNA editing of rpl16 in maize and Arabidopsis, respectively. Besides rpl16, EMP5 also affects the editing efficiency of some editing sites in other mitochondrial transcripts. Loss of function of EMP5 causes arrested embryogenesis and endosperm development in maize, but no discernable defective phenotype is observed in Arabidopsis otp72 mutant plants. We here report that EMP5 can completely complement Arabidopsis otp72 for Ararpl16-440 editing, and E domain can’t be disrupted for normal function of EMP5. An interesting finding was that overexpression of a set of truncated EMP5s in Col-0 resulted in delayed bolting phenotypes and smaller plants. It seems some truncated EMP5s, such as EMP5-C and EMP5-D, have yet unknown functions, are not responsible for Ararpl16-440 editing any more. The phylogenetic analysis indicated that the functions of most EMP5 orthologs may be conserved, but EMP5 ortholog proteins from Nicotiana and Solanum can’t be found, and the C to U editing of Zmrpl16-458 site was not needed any more in the two species. Therefore, these two primary orthologs maybe already evolve new functions and change to different new proteins during evolutionary history. Overall, our studies revealed that E domain is crucial for mitochondrial RNA editing function of EMP5, and a set of truncated EMP5s still have functions for Arabidopsis normal growth and development, providing new insights into investigating the functions of EMP5 protein.

As the most important major food crop, rice (Oryza sativa) often suffers from heat stress in tropical and subtropical regions. Thus, to investigate the mechanisms of responses and tolerance to heat stress (HS) at the miRNA level is important for breeding new rice varieties with HS tolerance. In this study, six miRNA libraries (with duplicates for each of the 0 h, 12 h and 24 h HS treatment) for HT54, an indica heat tolerant line, were sequenced. Based on this, a set of HS-responsive miRNAs and their target genes were identified by using the next-generation sequencing (NGS) and degradome sequencing (DS). A total of 419 conserved and 77 novel miRNAs were identified, of which 39 conserved and 2 novel miRNAs were found to be significantly differentially expressed (p < 0.05). As the most up-regulated miRNA, the expression of miR396 family members were verified by qRT-PCR. In addition, one down-regulated miR162b and one novel miRNA (pc-5p-93188_29) were experimentally validated. These results inferred that these loci were responsive to HS. Moreover, a total of 1364 target transcripts were identified for 114 conserved miRNAs and a total of 65 were identified for 22 novel miRNA families. GO and pathway enrichment analyses showed that these genes most likely play vital roles in rice HS tolerance. Taken together, a set of HS-responsive conserved and novel miRNAs were firstly characterized and identified in rice seedlings, which would lay a solid foundation for clarifying rice HS response and tolerance mechanisms.

As one of the most vital major food crops feeding almost half the earth's population, rice often suffers from heat stress in tropical and subtropical regions. To study rice heat stress tolerance mechanism is critical for breeding new heat tolerant rice varieties. We performed RNA-Seq analysis to check rice transcriptomic differences responsive to heat stress in both heat-tolerant (HT, HT54) and heat-sensitive (HS, HT13) lines. A sum of 0.603 billion reads were acquired on behalf of 32,391 unique genes. Compared with 0 h, a sum of 2275 genes showed enhanced regulation while 2270 genes showed inhibited regulation in the HS line at 12 h, contrarily 2603 genes showed enhanced regulation and 2900 genes showed inhibited regulation in the HT line. A sum of 1438 genes showed enhanced regulation and 1237 genes showed inhibited regulation in the HS line at 24 h, contrarily 1599 genes showed enhanced regulation and 1754 genes showed inhibited regulation in the HT line. About 22,490 genes were assigned to gene ontology (GO) term in terms of biological process, cellular component and molecular function categories. Compared with 0 h, a sum of 8 and 4 genes showed different expression (RFC ≥ 2.0) between the HT and HS rice lines at 12 and 24 h, respectively. This work most probably set a solid foundation for elucidating the heat stress tolerance mechanisms and crop genetic improvement in rice.

The presence of genetically modified (GM) protein in the endosperm is important information for the public when considering the biological safety of transgenic rice. To limit the expression of GM proteins to rice green tissues, we developed a modified Cre-lox gene switch using two cassettes named KEY and LOCK. KEY contains a nuclear-localized Cre recombinase driven by the green-tissue-specific promoter rbcS. LOCK contains a Nos terminator (NosT), which is used to block the expression of the gene of interest (GOI), bounded by two loxP sites. When KEY and LOCK are pyramided into hybrid rice, a complete gene switch system is formed. The Cre recombinase from KEY excises loxP-NosT in LOCK and unlocks the GOI in green tissues but keeps it locked in the endosperm. This regulatory effect was demonstrated by eYFP and Bt expression assays. The presence of eYFP and Cre were confirmed in the leaf, sheath, stem, and glume but not in the root, anther or seed of the gene-switch-controlled eYFP hybrids. Meanwhile, gene switch-controlled Bt hybrid rice not only confined the expression of Bt protein to the green tissues but also showed high resistance to striped stem borers and leaffolders.

Several mitochondrial-targeted pentatricopeptide repeat (PPR) proteins involved in pollen development have been reported to be fertility restorer (Rf) proteins. However, the roles of plastid-localized PPR proteins in plant male reproduction are poorly defined. Here, we described a plastid-localized PPR-SMR protein, OsPPR676, which is required for plant growth and pollen development in rice. In this study, OsPPR676 was confirmed to be an interacted protein with Osj10gBTF3, β-subunit of nascent polypeptide-associated complex (β-NAC), by bimolecular fluorescence complementation assays, indicating that both proteins are probably involved in the same regulatory pathway of pollen development. Compared with other chloroplast-rich tissues, OsPPR676 was only weakly expressed in anther, but in the Mei and YM stages of pollen development, its expression was relatively strong in the tapetum. Disruption of OsPPR676 resulted in growth retardation of plants and partial sterility of pollens. Phenotypic analysis of different osppr676 mutant lines implied that the SMR domain was not essential for the function of OsPPR676. We further demonstrated that OsPPR676 is essential for production of plastid atpB subunit, and then plays crucial roles in biosynthesis of fatty acids, carbohydrates, and other organic matters via affecting activity of ATP synthase.

Background: Lysine (Lys) is considered to be the first limiting essential amino acid in rice. Although there have been extensive efforts to improve the Lys content of rice through traditional breeding and genetic engineering, no satisfactory products have been achieved to date.Results: We expressed a LYSINE-RICH PROTEIN gene (LRP) from Psophocarpus tetragonolobus (L.) DC using an endosperm-specific GLUTELIN1 promoter (GT1) in Peiai64S (PA64S), an elite photoperiod-thermo sensitive male sterility (PTSMS) line. The expression of the foreign LRP protein was confirmed by Western blot analysis. The Lys level in the transgenic rice seeds increased more than 30 %, the total amount of other amino acids also increased compared to wild-type. Persistent investigation of amino acids in 3 generations showed that the Lys content was significantly increased in seeds of transgenic rice. Furthermore, Lys content in the hybrid of the transgenic plants also had an approximate 20 % increase compared to hybrid control. At the grain-filling stage, we monitored the transcript abundance of many genes encoding key enzymes involved in amino acid metabolism, and the results suggested that reduced amino acid catabolism led to the accumulation of amino acids in the transgenic plants. The genetically engineered rice showed unfavorable grain phenotypes compared to wild-type, however, its hybrid displayed little negative effects on grain.Conclusions: Endosperm-specific expression of foreign LRP significantly increased the Lys content in the seeds of transgenic plant, and the the Lys increase was stably heritable with 3 generation investigation. The hybrid of the transgenic plants also showed significant increases of Lys content in the seeds. These results indicated that expression of LRP in rice seeds may have promising applications in improving Lys levels in rice.

Iron (Fe) is an essential micronutrient for humans. Fe deficiency disease is widespread and has led to extensive studies on the mechanisms of Fe uptake and storage, especially in staple food crops such as rice. However, studies of functionally related genes in rice and other crops are often time and space demanding. Here, we demonstrate that transgenic Arabidopsis suspension culture cells and Arabidopsis plants can be used as an efficient expression system for gain-of-function study of selected transporters, using Fe transporters as a proof-of-principle. The vacuolar membrane transporters OsVIT1 and OsVIT2 have been described to be important for iron sequestration, and disruption of these two genes leads to Fe accumulation in rice seeds. In this study, we have taken advantage of the fluorescent-tagged protein GFP-OsVIT1, which functionally complements the Fe hypersensitivity of ccc1 yeast mutant, to generate transgenic Arabidopsis suspension cell lines and plants. GFP-OsVIT1 was shown to localize on the vacuolar membrane using confocal microscopy and immunogold EM. More importantly, the Fe concentration, as well as the concentration of Zn, in the transgenic cell lines and plants were significantly increased compared to that in the WT. Taken together, our study shows that the heterologous expression of rice vacuolar membrane transporter OsVIT1 in Arabidopsis system is functional and effectively enhances iron accumulation, indicating an useful approach for studying other putative transporters of crop plants in this system.

The identification result was consistent with the resistance evaluation to striped stem borer through four different bioassay methods, including detached-leaf bioassay, detached-stem bioassay, stripping detached-stem bioassay and whole plant bioassay. The detached-leaf bioassay is considered as the most rapid and effective method for evaluating resistance ability to striped stem borer. By both detached-leaf and whole plant bioassays, the author identified transgenic rice with Vip3A gene, and further validated the identi-fication effectiveness of the detached-leaf bioassay method. Simultaneously, our preliminary results revealed that detached-leaf bioas-say could also perform an accurate identification to rice leaf roller as lepidoptera pests of rice.

Eukaryotic translation initiation factor 3 (eIF3) is a large protein complex that participates in most translation initiation processes. While eIF3 has been well characterized, less is known about the roles of individual eIF3 subunits, particularly in plants. Here, we identified and characterized OseIF3e in rice (Oryza sativa L.). OseIF3e was constitutively expressed in various tissues, but most strongly in vigorously growing organs. Transgenic OseIF3e-silenced rice plants showed inhibited growth in seedling and vegetative stages. Repression of OseIF3e led to defects in pollen maturation but did not affect pollen mitosis. In rice, eIF3e interacted with eIF3 subunits b, d, e, f, h, and k, and with eIF6, forming homo- and heterodimers to initiate translation. Furthermore, OseIF3e was shown by yeast two-hybrid assay to specifically bind to inhibitors of cyclin-dependent kinases 1, 5, and 6. This interaction was mediated by the sequence of amino acid residues at positions 118-138, which included a conserved motif (IGPEQIETLYQFAKF). These results suggested although OseIF3e is not a "functional core" subunit of eIF3, it still plays crucial roles in rice growth and development, in combination with other factors. We proposed a pathway by which OseIF3e influence organ size and pollen maturation in rice, providing an opportunity to optimize plant architecture for crop breeding.

Gene pyramiding is an efficient approach for the genetic improvement of multiple agronomic traits simultaneously. In this study, we pyramided two foreign genes, cry1Ac driven by the rice Actin I promoter, and lysine-rich protein (LRP), driven by the endosperm-specific GLUTELIN1 (GT1) promoter, into the elite indica cultivar 9311. These two genes were chosen in an attempt to enhance insect-resistance and Lysine (Lys) content. In the pyramided line, the foreign gene cry1Ac was efficiently expressed in the leaves and stems, and exhibited highly efficient resistance to striped stem borer (SSB, Chilo suppressalis Walker) in the laboratory and rice leaf folder (RLF, Cnaphalocrocis medinalis Guenee) in the field. Furthermore, the LRP gene was highly expressed in the endosperm and produced a remarkable increase of Lys content in the seeds of the pyramided line. The data from field trials demonstrated that most of the agronomic traits including yield were well maintained in the pyramided line compared to the parental control. These results strongly suggest that the foreign cry1Ac and LRP genes have remarkable application potential in rice, and the resultant pyramided line serves as an ideal bridge material for the improvement of insect-resistance and high Lys rice in the future.

Tight and accurate regulation of immunity and thiamine biosynthesis is critical for proper defence mechanisms and several primary metabolic cycles in plants. Although thiamine is known to enhance plant defence by priming, the mechanism by which thiamine biosynthesis responds to immune signals remains poorly understood. Here we identified a novel rice (Oryza sativa L.) NB-LRR gene via an insertion mutation, this mutant confesses a low seed setting phenotype and the corresponding genetic locus was named OsLSR (Low seed setting related). Comparing with wildtype plant, both overexpression and suppression of OsLSR lead to the autoactivation of the rice immune system and accumulation of thiamine, which result in a great fitness cost and yield penalty. Moreover, when fused with eGFP at their C terminus, two fragments, OsLSR1-178 and OsLSR464-546, localized to chloroplasts where thiamine is produced. Our result suggests that OsLSR differs from traditional NB-LRR genes. Its expression is closely related to the immune status and thiamine level in plant cells and should be maintained within a narrow range for rice growth.