Dwarf Mutant Research Articles

Maize-Tripsacum-Teosinte allopolyploid (MTP), a novel dwarf mutant inducer tool in maize.

Dwarf plant architecture facilitates dense planting, and increased planting densities boost the maize yield. However, breeding applications of dwarfing materials for maize are currently limited. There is an urgent need remove the obstacles to applying dwarf resources. Here, we innovated a new method to add a novel maize dwarf germplasm through the distant hybridization of Maize-Tripsacum-Teosinte allopolyploid (MTP) with maize. We identified ten independent dwarf families with unique characteristics. Five germplasms in our library were controlled by their respective dwarf genes. However, no allele was controlled by Br2. Subsequently, d024 in the library was successfully fine mapped, revealing its linkage to indel-4 in ZmCYP90D1. The indel-4 polymorphism regulates the expression of ZmCYP90D1 and is controlled by an upstream transcription factor (ZmBES1/BZR1-5). The indel-4 of ZmCYP90D1 allele, which reduces plant height, originated from Tripsacum, a wild variety of maize. However, d024 exhibits sensitivity to brassinosteroids (BRs), with lower castasterone levels in the internodes than that in the wild type. Furthermore, ZmCYP90D1 interacted with ZmFDXs and ZmNAD(P)H to positively regulate the downstream BR synthesis pathway. Additionally, we showed that introgressing the indel-4 of the Tripsacum allele into modern hybrids ensures yield potential and improves the harvest index under high-density conditions. Overall, as we begin to manufacture highly engineered dwarf materials using the MTP, this approach will solve the problems faced by corn dwarfs.

A DUF21 domain-containing protein regulates plant dwarfing in watermelon.

Dwarf or semi-dwarf plant structures are well-suited for intensive farming, maximizing yield, and minimizing labor costs. Watermelon (Citrullus lanatus) is classified as an annual vine plant with elongated internodes, yet the mechanism governing watermelon dwarfing remains unclear. In this study, a compact watermelon mutant dwarf, induced by the insertion of T-DNA, was discovered. Through re-sequencing, a gene named domain of unknown function 21 (ClDUF21), located downstream of the T-DNA insertion site, was identified as the candidate gene for the dwarf mutant, and its functionality was subsequently confirmed. Watermelon mutants generated through CRISPR/Cas9-mediated knockout of ClDUF21 revealed that homozygous mutants displayed a pronounced dwarfing phenotype, and protein-protein interaction analysis confirmed the direct interaction between ClDUF21 and ClDWF1. Subsequently, we employed CRISPR/Cas9 technology to precisely modify the homologous gene CsDUF21 in cucumber (Cucumis sativus) and performed protein interaction validation between CsDUF21 and CsDWF1, thereby demonstrating that the CsDUF21 gene also exhibits analogous functionality in plant dwarfing. These findings demonstrate that ClDUF21 governs plant dwarfism by modulating the brassinosteroid synthesis pathway via ClDWF1.

Genome-Wide Identification of Height-Related Genes Using Three Maize Dwarfs and RNA-Seq

Plant height is an important grain yield-associated trait in maize. To date, few genes related to plant height have been characterized in maize. To better understand the genetic mechanisms of plant height in maize, we revealed the transcriptional changes of three dwarf mutants compared to the wild type. By ethyl methane sulfonate treatment of the wild-type maize cultivar PH6WC, we obtained three dwarfs—PH6WCdwarf1 (pd1), PH6WCdwarf2 (pd2), and PH6WCdwarf3 (pd3)—and their plant heights were reduced by 42%, 38%, and 24%, respectively. RNA-Seq data suggested that 1641 differentially expressed genes (DEGs) overlapped with each other among the three dwarfs at the seedling stage. Further analysis showed that the DEGs were divided into four groups with different expression patterns. Functional analysis revealed that these DEGs were commonly enriched in 47 GO terms mainly involved in cytokinesis, hormone, and energy metabolism pathways. Among them, An1, involved in the GA biosynthesis pathway, and mutations in An1 result in reduced plant height. EREB182 encodes ethylene-responsive element binding protein 2, which is critical for internode elongation. Microtubule-related genes Zmtub2, Zmtub3, Zmtub5, Zmtub6, and TUBG2 were commonly enriched among the three comparisons. Previous studies have shown that mutations in microtubule-associated genes cause the dwarf phenotype. However, nearly half of the common DEGs had no functional information, such as Zm00001d000107, Zm00001d000279, etc., implying their novel and specific functions in maize. Overall, this study identifies several potential plant height-related genes and contributes to linking genetic resources with maize breeding.

Comparative Analysis of Alternative Splicing in Moso Bamboo and Its Dwarf Mutant, Phyllostachys edulis ‘Tubaeformis’

Internode length is a crucial phenotypic trait of bamboo, significantly impacting its processing and utilization. Phyllostachys edulis ‘Tubaeformis’ (Shengyin Bamboo), a variety of Moso bamboo, exhibits drastically shortened internodes, making it a valuable ornamental bamboo species. We used PacBio single-molecule long-read sequencing and second-generation sequencing to identify genome-wide alternative splicing (AS) events in Moso bamboo and its dwarf mutant, Shengyin bamboo, and compared the differences between the two. Our sequencing data unveiled 139,539 AS events, with retained introns as the most prevalent events. A large number of genes were differentially alternatively spliced (DAS) between Moso bamboo and Shengyin bamboo, and genes related to RNA splicing were most significantly enriched. The high expression of SR isoforms in the 24th internode of Moso bamboo is likely the main factor leading to its greater number of alternative splicing events. Alternative splicing affects the functional domains of partial GRF, E2F, and NAM isoforms, leading to the loss of domains in some isoforms and enabling some isoforms to acquire new functional domains, and this phenomenon is more common in Shengyin bamboo. AS modifies the functional domains of certain GRF isoforms, frequently resulting in domain losses or endowing isoforms with novel domains, and this phenomenon is more common in Shengyin bamboo. We used PacBio single-molecule long-read sequencing and second-generation sequencing to identify genome-wide alternative splicing (AS) events in Moso bamboo and its dwarf mutant, Shengyin Bamboo and compared the differences between the two.

Identification and characterization of CsERECTA, a major gene controlling stem elongation through regulating GA biosynthesis in cucumber.

Dwarfing is an ideal agronomic trait in crop breeding, which can improve lodging resistance and increase crop productivity. In this study, we identified a dwarf mutant cp-3 from an EMS-mutagenized population, which had extremely short internodes, and the cell length and number of internodes were significantly reduced. Meanwhile, exogenous GA3 treatment partially rescued the plant height of the cp-3. Inheritance analysis showed that the cp-3 mutant was regulated via a recessive nuclear locus. A candidate gene, CsERECTA, encoding an LRR receptor-like serine/threonine-protein kinase, was cloned through a map-based cloning strategy. Sequence analysis showed that a nucleotide mutation (C ~ T) in exon 26 of CsERECTA led to premature termination of the protein. Subsequently, two transgenic lines were generated using the CRISPR/Cas9 system, and they showed plant dwarfing. Plant endogenous hormones quantitative and RNA-sequencing analysis revealed that GA3 content and the expression levels of genes related to GA biosynthesis were significantly reduced in Cser knockout mutants. Meanwhile, exogenous GA3 treatment partially rescued the dwarf phenotype of Cser knockout mutants. These findings revealed that CsERECTA controls stem elongation by regulating GA biosynthesis in cucumber.

Overexpression of Calcineurin B-like Interacting Protein Kinase 31 Promotes Lodging and Sheath Blight Resistance in Rice.

A breakthrough "Green Revolution" in rice enhanced lodging resistance by using gibberellin-deficient semi-dwarf varieties. However, the gibberellic acid (GA) signaling regulation on rice disease resistance remains unclear. The resistance test showed that a positive GA signaling regulator DWARF1 mutant d1 was more susceptible while a negative GA signaling regulator Slender rice 1 (SLR1) mutant was less susceptible to sheath blight (ShB), one of the major rice diseases, suggesting that GA signaling positively regulates ShB resistance. To isolate the regulator, which simultaneously regulates rice lodging and ShB resistance, SLR1 interactors were isolated. Yeast two-hybrid (Y2H), bimolecular fluorescence complementation (BiFC), and Co-IP assay results indicate that SLR1 interacts with Calcineurin B-like-interacting protein kinase 31 (CIPK31). cipk31 mutants exhibited normal plant height, but CIPK31 OXs showed semi-dwarfism. In addition, the SLR1 level was much higher in CIPK31 OXs than in the wild-type, suggesting that CIPK31 OX might accumulate SLR1 to inhibit GA signaling and thus regulate its semi-dwarfism. Recently, we demonstrated that CIPK31 interacts and inhibits Catalase C (CatC) to accumulate ROS, which promotes rice disease resistance. Interestingly, CIPK31 interacts with Vascular Plant One Zinc Finger 2 (VOZ2) in the nucleus, and expression of CIPK31 accumulated VOZ2. Inoculation of Rhizoctonia solani AG1-IA revealed that the voz2 mutant was more susceptible to ShB. Thus, these data prove that CIPK31 promotes lodging and ShB resistance by regulating GA signaling and VOZ2 in rice. This study provides a valuable reference for rice ShB-resistant breeding.

Low mutation rate of spontaneous mutants enables detection of causative genes by comparing whole genome sequences.

In the early 1900s, mutation breeding to select varieties with desirable traits using spontaneous mutation was actively conducted around the world, including Japan. In rice, the number of fixed mutations per generation was estimated to be 1.38-2.25. Although this low mutation rate was a major problem for breeding in those days, in the modern era with the development of next-generation sequencing (NGS) technology, it was conversely considered to be an advantage for efficient gene identification. In this paper, we proposed an in silico approach using NGS to compare the whole genome sequence of a spontaneous mutant with that of a closely related strain with a nearly identical genome, to find polymorphisms that differ between them, and to identify the causal gene by predicting the functional variation of the gene caused by the polymorphism. Using this approach, we found four causal genes for the dwarf mutation, the round shape grain mutation and the awnless mutation. Three of these genes were the same as those previously reported, but one was a novel gene involved in awn formation. The novel gene was isolated from Bozu-Aikoku, a mutant of Aikoku with the awnless trait, in which nine polymorphisms were predicted to alter gene function by their whole-genome comparison. Based on the information on gene function and tissue-specific expression patterns of these candidate genes, Os03g0115700/LOC_Os03g02460, annotated as a short-chain dehydrogenase/reductase SDR family protein, is most likely to be involved in the awnless mutation. Indeed, complementation tests by transformation showed that it is involved in awn formation. Thus, this method is an effective way to accelerate genome breeding of various crop species by enabling the identification of useful genes that can be used for crop breeding with minimal effort for NGS analysis.

Cloning and functional mechanism of the dwarf gene gba affecting stem elongation and cellulose biosynthesis in jute (Corchorus olitorius).

Plant height (PH) is an important factor affecting bast fiber yield in jute. Here, we report the mechanism of dwarfism in the 'Guangbaai' (gba) of jute. The mutant gba had shorter internode length and cell length compared to the standard cultivar 'TaiZi 4' (TZ4). Exogenous GA3 treatment indicated that gba is a GA-insensitive dwarf mutant. Quantitative trait locus (QTL) analysis of three PH-related traits via a high-density genetic linkage map according to re-seq showed that a total of 25 QTLs were identified, including 13 QTLs for PH, with phenotypic variation explained ranging from 2.42 to 74.16%. Notably, the functional mechanism of the candidate gene CoGID1a, the gibberellic acid receptor, of the major locus qPHIL5 was evaluated by transgenic analysis and virus-induced gene silencing. A dwarf phenotype-related single nucleotide mutation in CoGID1a was identified in gba, which was also unique to the dwarf phenotype of gba among 57 cultivars. Cogid1a was unable to interact with the growth-repressor DELLA even in the presence of highly accumulated gibberellins in gba. Differentially expressed genes between transcriptomes of gba and TZ4 after GA3 treatment indicated up-regulation of genes involved in gibberellin and cellulose synthesis in gba. Interestingly, it was found that up-regulation of CoMYB46, a key transcription factor in the secondary cell wall, by the highly accumulated gibberellins in gba promoted the expression of cellulose synthase genes CoCesA4 and CoCesA7. These findings provide valuable insights into fiber development affected by endogenous gibberellin accumulation in plants.

A mutation in the brassinosteroid biosynthesis gene CpDWF5 disrupts vegetative and reproductive development and the salt stress response in squash (Cucurbita pepo).

A Cucurbita pepo mutant with multiple defects in growth and development has been identified and characterized. The mutant dwfcp displayed a dwarf phenotype with dark green and shrinking leaves, shortened internodes and petioles, shorter but thicker roots and greater root biomass, and reduced fertility. The causal mutation of the phenotype was found to disrupt gene Cp4.1LG17g04540, the squash orthologue of the Arabidopsis brassinosteroid (BR) biosynthesis gene DWF5, encoding for 7-dehydrocholesterol reductase. A single nucleotide transition (G > A) causes a splicing defect in intron 6 that leads to a premature stop codon and a truncated CpDWF5 protein. The mutation co-segregated with the dwarf phenotype in a large BC1S1 segregating population. The reduced expression of CpDWF5 and brassinolide (BL) content in most mutant organs, and partial rescue of the mutant phenotype by exogenous application of BL, showed that the primary cause of the dwarfism in dwfcp is a BR deficiency. The results showed that in C. pepo, CpDWF5 is not only a positive growth regulator of different plant organs but also a negative regulator of salt tolerance. During germination and the early stages of seedling development, the dwarf mutant was less affected by salt stress than the wild type, concomitantly with a greater upregulation of genes associated with salt tolerance, including those involved in abscisic acid (ABA) biosynthesis, ABA and Ca2+ signaling, and those coding for cation exchangers and transporters.

Molecular and physiological characterization of early semi-dwarf mutants of rice and localization of SNP variants in Sd1 locus generated through gamma radiation

Purpose The ‘Improved White Ponni’ (IWP) rice variety, which is susceptible to lodging, leading to yield losses. Our primary goal is to develop new rice lines with non-lodging traits, enhancing stem strength and resistance to adverse conditions. Additionally, we aim to improve yield-contributing agronomic traits, benefiting farmers, food security, and the environment. Our work contributes to scientific knowledge and addresses a significant issue in Southern Indian rice cultivation. Materials and Methods In the present study, early and semi-dwarf early mutants of IWP were developed without altering the native grain quality traits using gamma ray-mediated mutagenesis. The seeds (500) were irradiated with γ-rays after fixing the Lethal Dose 50 (LD50), and selection for semi-dwarfism and earliness was imposed on a large M2 population. The selected traits were confirmed by evaluating the M3 lines at morpho-physiological, biochemical, and molecular levels. Results The response of mutants to gibberellic acid has been studied, which identified responsive mutants as well as slow-responding mutant lines including IWP-11-2, IWP-48-2, IWP-50-11, and IWP-33-2. Agar plate assay indicated low α- amylase content in IWP-50-11, IWP-33-2, IWP-43-1, IWP-47-2, and IWP-18-1. The scanning electron microscopy demonstrated that the mutants displayed an increased cellular dimension in comparison to the wild type. In dwarf mutants, null alleles were observed for the SD1 gene-specific primers which depicts gene undergone mutation. Further sequencing revealed the presence of single nucleotide polymorphisms in the SD1 gene resulting in semi-dwarfism in the mutant IWP-D-1. Conclusions The impact of a defective gibberellic acid-mediated signaling pathway in mutants to produce a novel high-yielding and early maturing semi-dwarf rice variety.

A mutation in CsDWF7 gene encoding a delta7 sterol C-5(6) desaturase leads to the phenotype of super compact in cucumber (Cucumis sativus L.).

A novel super compact mutant, scp-3, was identified using map-based cloning in cucumber. The CsDWF7 gene encoding a delta7 sterol C-5(6) desaturase was the candidate gene of scp-3. Mining dwarf genes is important in understanding stem growth in crops. However, only a small number of dwarf genes have been cloned or characterized. Here, we characterized a cucumber (Cucumis sativus L.) dwarf mutant, super compact 3 (scp-3), which displays shortened internodes and dark green leaves with a wrinkled appearance. The photosynthetic rate of scp-3 is significantly lower than that of the wild type. The dwarf phenotype of scp-3 mutant can be partially rescued by the exogenous brassinolide (BL) application, and the endogenous brassinosteroids (BRs) levels in the scp-3 mutant were significantly lower compared to the wild type. Microscopic examination revealed that the reduced internode length in scp-3 resulted from a decrease in cell size. Genetic analysis showed that the dwarf phenotype of scp-3 was controlled by a single recessive gene. Combined with bulked segregant analysis and map-based cloning strategy, we delimited scp-3 locus into an 82.5kb region harboring five putative genes, but only one non-synonymous mutation (A to T) was discovered between the mutant and its wild type in this region. This mutation occurred within the second exon of the CsGy4G017510 gene, leading to an amino acid alteration from Leu156 to His156. This gene encodes the CsDWF7 protein, an analog of the Arabidopsis DWF7 protein, which is known to be involved in the biosynthesis of BRs. The CsDWF7 protein was targeted to the cell membrane. In comparison to the wild type, scp-3 exhibited reduced CsDWF7 expression in different tissues. These findings imply that CsDWF7 is essential for both BR biosynthesis as well as growth and development of cucumber plants.

DWARF AND LOW-TILLERING 2 functions in brassinosteroid signaling and controls plant architecture and grain size in rice.

Brassinosteroids (BRs) are a class of steroid phytohormones that control various aspects of plant growth and development. Several transcriptional factors (TFs) have been suggested to play roles in BR signaling. However, their possible relationship remains largely unknown. Here, we identified a rice mutant dwarf and low-tillering 2 (dlt2) with altered plant architecture, increased grain width, and reduced BR sensitivity. DLT2 encodes a GIBBERELLIN INSENSITIVE (GAI)-REPRESSOR OF GAI (RGA)-SCARECROW (GRAS) TF that is mainly localized in the nucleus and has weak transcriptional activity. Our further genetic and biochemical analyses indicate that DLT2 interacts with two BR-signaling-related TFs, DLT and BRASSINAZOLE-RESISTANT 1, and probably modulates their transcriptional activity. These findings imply that DLT2 is implicated in a potentially transcriptional complex that mediates BR signaling and rice development and suggests that DLT2 could be a potential target for improving rice architecture and grain morphology. This work also sheds light on the role of rice GRAS members in regulating numerous developmental processes.

Phenotypic Characterization and Gene Mapping of a Spiral Leaf and Dwarf (sld) Mutant from Tetraploid Common Tobacco (Nicotiana tabacum L.)

Leaf morphology and plant height are two agronomic traits closely related to tobacco (Nicotiana tabacum L.) yield and quality. The study of leaf morphology and plant stature mutants will greatly contribute to the fields of plant architecture breeding and developmental biology. Here, we report the characterization of a spiral leaf and dwarf (sld) mutant identified from an ethylmethane sulfonate (EMS)-induced common tobacco population. The sld mutant displayed the phenotype of wrinkled, spiral, and miniature leaves, with the growth point as the central axis and plant dwarfing with shortened internodes. The inheritance pattern of the sld mutant phenotype was manipulated by a recessive nuclear monogene, which was linked to six tobacco simple sequence repeat (SSR) markers from linkage group 5 via gene mapping. Utilizing an F2 population, the sld mutant gene the sld mutant gene was located between the co-segregated markers PT51778, PT54913, and the marker PT61414, with an equal genetic distance of 0.16 cM. Taking advantage of a BC1F1 population, the markers PT51778, PT54913, the sld gene, and the marker PT61414 demonstrated co-segregation, located between the markers PT40040 and PT60933, respectively, with a genetic distance of 1.37 cM and 6.32 cM, respectively. These findings will be helpful in cloning the sld gene and in the further characterization of the regulatory genes controlling the spiral and dwarfing phenotypes in tobacco.

Inheritance of dwarfism trait by winter wheat mutants induced in the Chornobyl NPP exclusion zone

Background. The development of short-stem cultivars is an effective way to increase the productivity of winter wheat grown under the intensive cultivation techno­logy. Dwarf mutants, induced in the exclusion zone of ChNPP, can preserve a productive potential of an initial cultivar and be of a breeding value. The knowledge of the inheri­tance regularities of stem length by dwarf winter wheat mutants makes it possible to choose the pairs for crossing better and predict a final result in hybrid generations earlier. Materials and methods. Hybrids F1 і F2 of Triticum aestivum L. were received by crossing medium-grown cultivar Sonechko with dwarf mutant lines UK 1145/10, UK 1147/10, and UK 1148/10, induced by the effect of the radionuclide contamination of the exclusion zone of ChNNP on the plants of ‘Albatros odeskyi’ cultivar. The nature of the trait inheritance in F1 was determined by the degree of phenotype dominance, the variability of plant height was determined by a variation coefficient. The segregation frequency of dwarf, low-, medium- and high-grown forms was accounted in the population of F2 plants. Results. By stem length, plants F1 exceeded the genotypes involved in hybridization, or were closer to parental forms with a greater manifestation of the trait. The real heterosis in the plant height decrease was shown in the F1 hybrid ‘Sonechko’ × UK 1147/10. The hybrid productivity elements are inherited by the type of a partial positive dominance and over-dominance. From 2.2 % to 3.6 % of high-grown forms, atypical for parents, were recorded in the population of F2. Conclusions. When the crossing combination includes a medium-grown cultivar of winter wheat and dwarf mutants, induced in the exclusion zone of ChNPP, a stem length in F1 is inherited by an intermediate type and partial positive dominance. Dwarf mutant UK 1147/10 can be valuable for breeding short stem winter bread wheat. A significant variation of such indicator as a degree of phenotype dominance by all studied features confirms a complicated nature of genetic determination of a plant height and the productivity elements of winter wheat, in the formation of which various types of gene interaction take part.

Induction and Characterization of Mutations Related to Dwarf Habit in Hardy Hibiscus (Muenchhusia section)

Hybrids between various species of Hibiscus in the Muenchhusia section are popular in commercial markets due to their colorful, floriferous blooms. However, many commercial cultivars are too vigorous for use in smaller garden spaces and often have only a few flowers in bloom. Stable, heritable mutants exhibiting dwarf stature and better branching architecture will be valuable to the commercial production of hardy Hibiscus. The goal of this project was to induce stable, heritable mutations using ethyl methanesulfonate that result in increased branching and a dwarf compact phenotype and introgress the compact phenotypes into lines of hardy Hibiscus to generate a series of diverse compact cultivars for use in commercial markets. Multiple mutations associated with dwarf, compact phenotypes were successfully induced. Hybridizations were made between M2 plants displaying a dwarf phenotype and hybrids developed through conventional breeding that do not possess the dwarf mutation in their background to develop diverse selections of dwarf plants. Inheritance patterns of the dwarf genes were determined through the segregation ratios of the dwarf phenotype in the F1 and F2 generations of these hybrids. Compared with wild-type progeny, dwarf progeny had shorter internode lengths and a greater number of primary branches.

Gibberellin metabolism and signaling.

Gibberellins (GAs) are plant hormones with a tetracyclic diterpenoid structure that are involved in various important developmental processes. Two GA-deficient mutants were isolated: a semidwarf mutant "sd1", which was found to have a defective GA20ox2 gene and was introduced to the world in a green revolution cultivar, and a severe dwarf allele of "d18", with a defective GA3ox2 gene. Based on the phenotypic similarity of d18, rice dwarf mutants were screened, further classifying them into GA-sensitive and GA-insensitive by applying exogenous GA3. Finally, GA-deficient rice mutants at 6 different loci and 3 GA signaling mutants (gid1, gid2, and slr1) were isolated. The GID1 gene encodes a GA nuclear receptor, and the GID1-DELLA (SLR1) system for GA perception is widely used in vascular plants. The structural characteristics of GID1 and GA metabolic enzymes have also been reviewed.

Brassinosteroid coordinates cell layer interactions in plants via cell wall and tissue mechanics.

Growth coordination between cell layers is essential for development of most multicellular organisms. Coordination may be mediated by molecular signaling and/or mechanical connectivity between cells, but how genes modify mechanical interactions between layers is unknown. Here we show that genes driving brassinosteroid synthesis promote growth of internal tissue, at least in part, by reducing mechanical epidermal constraint. We identified a brassinosteroid-deficient dwarf mutant in the aquatic plant Utricularia gibba with twisted internal tissue, likely caused by mechanical constraint from a slow-growing epidermis. We tested this hypothesis by showing that a brassinosteroid mutant in Arabidopsis enhances epidermal crack formation, indicative of increased tissue stress. We propose that by remodeling cell walls, brassinosteroids reduce epidermal constraint, showing how genes can control growth coordination between layers by means of mechanics.

Transcriptome analysis for deep understanding the dwarfing mechanism of Malus dwarf mutant rootstock A1d

A1d is dwarf mutant of Pingyi Tiancha (M. hupehensis var. Pinyiensis, PYTC) with dwarfing effect comparable to that of M26. In order to understanding how it affected scion shoot growth at transcriptional level and identify potential useful dwarf genes from it, the transcriptomic differences between the new spring shoots of apple variety “Fuji6” grafted on vigorous rootstock Pingyi Tiancha (P-F) and A1d (A-F) were investigated by RNA-Seq and followed by qRT-PCR verification in two types of scions as well as rootstocks. 494 differentially expressed genes (DEGs) were identified between two samples by RNA-Seq. Plant signal transduction, phytohormone metabolism and IAA polar transportation pathways were greatly affected by the dwarf A1d rootstock. In particular, gibberellin 2-beta-dioxygenase (GA2OX), which encodes a gibberellic acid (GA) inactivating gene, was down-regulated both in A1d rootstock and A-F, while the contents of active GA forms were higher in A-F shoots. The results of qRT-PCR revealed that genes related with IAA signal, IAA polar transportation and cell wall degradation etc. were significantly down-regulated A1d rootstock and its scions. However, down-regulation of these genes in A-F was not retrieved by GA treatment. Sequence analysis showed that two amino acids in the conserved N-terminal DELLA region of Malus domestica RGA-like 3b (MhRGL3b) were mutated in one chromosome of A1d, and designated as Mhrgl3b. We propose that the transportation of the Mhrgl3b from A1d rootstock play a dominant role in controlling the vigor of scion.

Exogenous hormone application regulates dwarf mutant plant height in Sophora davidii (Franch.) Skeels by changing endogenous hormone levels

Dwarf mutants are valuable materials for investigating plant gene function and developing new crop varieties. In the present study, dwarf mutant (D-CK) and wild-type (W-CK) Sophora davidii (Franch.) Skeels plants foliarly sprayed with distilled water were used as controls, and dwarf mutants were treated with different concentrations of gibberellin A3 (GA3; 20, 40 and 60 mg L-1), brassinosteroid (BR; 12.5, 25 and 50 mg L-1); and auxin indole-3-acetic acid (IAA; 12.5, 25 and 50 mg L-1) to study the changes in plant height at 20, 40 and 60 d after exogenous hormone spraying. The changes in endogenous contents of IAA, GA3, BR, abscisic acid (ABA), jasmonic acid (JA), and salicylic acid (SA) and IAA:ABA ratio in the dwarf mutant plants were also measured at 20, 40 and 60 d after exogenous hormone application. The dwarf mutants reverted to the W-CK height phenotype after 60 d of exogenous spraying with different concentrations of GA3 and BR (except 50 mg·L-1), but after exogenous spraying with IAA, dwarf mutants did not revert to the W-CK height phenotype. Compared with D-CK plants, endogenous GA3, BR and IAA contents in dwarf mutant plants increased to some extent with exogenous spraying of GA3, BR and IAA. The results indicated that exogenous spraying of different concentrations of GA3 and BR regulated S. davidii plant height by promoting the secretion of endogenous GA3, BR, IAA and SA and inhibiting endogenous ABA secretion in dwarf mutant. The height of mutants was sensitive to exogenous GA3 and BR but not to exogenous IAA.

Fine Mapping of the Affecting Tillering and Plant Height Gene CHA-1 in Rice

The plant architecture of rice is an important factor affecting yield. Strigolactones (SLs) are newly discovered carotenoid-derived plant hormones that play an important role in rice plant architecture. In this study, a high-tillering dwarf mutant, CHA-1, was identified by spatial mutagenesis. CHA-1 was located in the region of 31.52-31.55 MB on chromosome 1 by map-based cloning. Compared with the wild-type THZ, the CHA-1 mutant showed that ACCAC replaced TGGT in the coding region of the candidate gene LOC_Os01g54810, leading to premature termination of expression. Genetic complementation experiments proved that LOC_Os01g54810 was CHA-1, which encodes a putative member of Class III lipase. Expression analysis showed that CHA-1 was constitutively expressed in various organs of rice. Compared with those in THZ, the expression levels of the D17 and D10 genes were significantly downregulated in the CHA-1 mutant. In addition, the concentrations of ent-2'-epi-5-deoxystrigol (epi-5DS) in the root exudates of the CHA-1 mutant was significantly reduced compared with that of THZ, and exogenous application of GR24 inhibited the tillering of the CHA-1 mutant. These results suggest that CHA-1 influences rice architecture by affecting SL biosynthesis.