A Heterozygous Genotype-Dependent Branched-Spike Wheat and the Potential Genetic Mechanism Revealed by Transcriptome Sequencing

Wheat (Triticum aestivum L.) spike architecture is an important trait associated with spike development and grain yield. Here, we report a naturally occurring wheat mutant with branched spikelets (BSL) from its wild-type YD-16, which has a normal spike trait and confers a moderate level of resistance to wheat Fusarium head blight (FHB). The lateral meristems positioned at the basal parts of the rachis node of the BSL mutant develop into ramified spikelets characterized as multiple spikelets. The BSL mutant shows three to four-day longer growth period but less 1000-grain weight than the wild type, and it becomes highly susceptible to FHB infection, indicating that the locus controlling the BSL trait may have undergone an intensively artificial and/or natural selection in modern breeding process. The self-pollinated descendants of the lines with the BSL trait consistently segregated with an equal ratio of branched and normal spikelets (NSL) wheat, and homozygotes with the BSL trait could not be achieved even after nine cycles of self-pollination. Distinct segregation patterns both from the self-pollinated progenies of the BSL plants and from the reciprocal crosses between the BSL plants with their sister NSL plants suggested that gametophytic male sterility was probably associated with the heterozygosity for the BSL trait. Transcriptome sequencing of the RNA bulks contrasting in the two types of spike trait at the heading stage indicated that the genes on chromosome 2DS may be critical for the BSL trait formation since 329 out of 2540 differentially expressed genes (DEGs) were located on that chromosome, and most of them were down-regulated. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that carbohydrate metabolism may be involved in the BSL trait expression. This work provides valuable clues into understanding development and domestication of wheat spike as well as the association of the BSL trait with FHB susceptibility.

Genome-Wide Identification and Characterization of wALOG Family Genes Involved in Branch Meristem Development of Branching Head Wheat

The branched spike phenotype is an important supernumerary spikelet trait of Triticum turgidum L. associated with the production of significantly more grains per spike, thereby offering a higher potential yield. However, the genetic basis of branch meristem (BM) development remains to be fully elucidated in wheat. TAW1, an ALOG (Arabidopsis LSH1 and Oryza G1) family gene, has been shown to function as a unique regulator in promoting BM development in rice. In this study, we found that the development pattern of the BMs of the branched spike in wheat was similar to the indeterminate BMs of rice. Moreover, phylogenetic analysis classified the ALOG genes into 12 groups. This family of genes was found to have evolved independently in eudicots and monocots and was evolutionarily conserved between wheat and rice as well as during wheat polyploidization. Furthermore, experiments revealed that TtALOG2-1A, a TAW1-homologous gene, plays a significant role in regulating the transition of indeterminate BM fate. Finally, large-scale RNA-sequencing studies and quantitative real-time polymerase chain reaction (qRT-PCR) experiments revealed that members of the TtALOGs may act upstream of the TtMADS22, TtMADS47, and TtMADS55 genes to promote indeterminate BM activities. Our findings further knowledge on BM development in wheat.

Auxins Regulations of Branched Spike Development and Expression of TFL, a LEAFY-Like Gene in Branched Spike Wheat (Triticum aestivum)

Branched spike wheat is a hexaploid germplasm with branched rachis on its main rachises, and the crucial period for branched rachises occurrence and development is just after the two ridges stage of shoot apex. Natural [indole-3-acetic acid (IAA), indole-3butyric acid (IBA)] and synthetic [(1-naphthaleneacetic acid (NAA), 2,4-Dichlorophenoxyacetic acid (2,4-D)] auxins were applied at this period to investigate the spike traits, seedling growth and photosynthesis related characters and expression of a putative homologue of the LEAFY in branched spike wheat. The four types of experienced auxins induced similar effects on these foresaid characters, although the impact extents were different among the auxins treatments. More branched rachis, spikelets, fertile florets and longer branched rachis were obtained in plants with IAA and IBA at 0.1 mM or NAA and 2,4-D at 1.0mM than those plants with no auxin treated. Auxin treatments also increased fresh and dry mass, photosynthetic pigment and parameters. TFL, a LEAFY-like gene was cloned in branched spike wheat and TFL mRNA expression was quantified using real-time reverse transcriptase-PCR. Application of the auxins accelerated the rise in TFL expression during the periods of branched rachises occurrence and extension. The data supports the hypothesis that auxins play a central role in the regulation branched spike development and TFL might correlate with the development of branched rachises in branched spike wheat.