Gene regulation network analyses of pistil development in papaya

Background The pistil is an essential part of flowers that functions in the differentiation of the sexes and reproduction in plants. The stigma on the pistil can accept pollen to allow fertilization and seed development. Papaya (Carica papaya L.) is a dioecious plant, where female flowers exhibit normal pistil, while the male flowers exhibit aborted pistil at a late stage of pistil development. Results The developmental stages of papaya pistil were analyzed after first dividing it into slices representing the primordium stage 1 (S1), the pre-meiotic stages S2, post-meiotic stage S3, and the mitotic stage S4. The SS scoring algorithm analysis of genes preferentially expressed at different stages revealed differentially expressed genes between male and female flowers. A transcription factor regulatory network for each stage based on the genes that are differentially expressed between male and female flowers was constructed. Some transcription factors related to pistil development were revealed based on the analysis of regulatory networks such as CpAGL11, CpHEC2, and CpSUPL. Based on the specific expression of genes, constructed a gene regulatory subnetwork with CpAGL11-CpSUPL-CpHEC2 functioning as the core. Analysis of the functionally enriched terms in this network reveals several differentially expressed genes related to auxin/ brassinosteroid signal transduction in the plant hormone signal transduction pathway. At the same time, significant differences in the expression of auxin and brassinosteroid synthesis-related genes between male and female flowers at different developmental stages were detected. Conclusions The pistil abortion of papaya might be caused by the lack of expression or decreased expression of some transcription factors and hormone-related genes, affecting hormone signal transduction or hormone biosynthesis. Analysis of aborted and normally developing pistil in papaya provided new insights into the molecular mechanism of pistil development and sex differentiation in dioecious papaya.

Lecciana, a New Low-Vigour Olive Cultivar Suitable for Super High Density Orchards and for Nutraceutical EVOO Production

Cultivar is the key factor for sustainability of the olive super high density planting system (SHD). ‘Lecciana’ is a new olive cultivar for oil production obtained in 1998 by a controlled cross between cv. Arbosana (♀) and cv. Leccino (♂) in a breeding program as part of an international research agreement between Agromillora Iberia S.L.U. and University of Bari. ‘Lecciana’ is the first olive cultivar of Italian descent suitable for SHD, featuring all the vegetative and productive traits required for efficient, sustainable olive growing intensification. Thanks to low vigor, early bearing (3rd year after planting), high yield efficiency (about 0.5 kg of fruits cm−2 of trunk section area) and good fruit size (3.5 g), ‘Lecciana’ could be planted with tree densities over 1,200 trees per hectare for an efficient continuous mechanical harvesting. High frost resistance, very low pistil abortion (3%), high fruit set (3%), oil content (over 19% fw) and, above all, good unsaturated fatty acids profile, polyphenols content (over 450 mg kg−1) and fruitiness median are the main distinctive characters of this new cultivar. The oils of ‘Lecciana’ fall into the category ‘nutraceutical EVOOs’ which can benefit from the specific functional health claim.

Phytohormone and integrated mRNA and miRNA transcriptome analyses and differentiation of male between hermaphroditic floral buds of andromonoecious Diospyros kaki Thunb

Background Persimmon (Diospyros kaki Thunb.) has various labile sex types, and studying its sex differentiation can improve breeding efficiency. However, studies on sexual regulation patterns in persimmon have focused mainly on monoecy and dioecy, whereas little research has been published on andromonoecy. In order to reveal the sex differentiation regulation mechanism of andromonoecious persimmon, we performed histological and cytological observations, evaluated OGI and MeGI expression and conducted phytohormones assays and mRNA and small RNA transcriptome analyses of the male and hermaphroditic floral buds of the andromonoecious persimmon ‘Longyanyeshi 1’. Results Stages 2 and 4 were identified as the critical morphological periods for sex differentiation of ‘Longyanyeshi 1’ by histological and cytological observation. At both stages, OGI was differentially expressed in male and hermaphroditic buds, but MeGI was not. This was different from their expressions in dioecious and monoecious persimmons. Meantime, the results of phytohormones assays showed that high IAA, ABA, GA3, and JA levels at stage 2 may have promoted male floral bud differentiation. However, high JA levels at stage 4 and high ZT levels at stages 2 and 4 may have promoted hermaphroditic floral bud differentiation. In these phytohormone biosynthesis and signaling pathways, 52 and 54 differential expression genes (including Aux/IAA, ARFs, DELLA, AHP, A-ARR, B-ARR, CYP735A, CRE1, PP2C, JAZ, MYC2, COI1, CTR1, SIMKK, ACO, and MPK6) were identified, respectively. During the development of male floral buds, five metacaspases genes may have been involved in pistil abortion. In addition, MYB, FAR1, bHLH, WRKY, and MADS transcription factors might play important roles in persimmon floral bud sex differentiation. Noteworthy, miR169v_1, miR169e_3, miR319_1, and miR319 were predicted to contribute to phytohormone biosynthesis and signaling pathways and floral organogenesis and may also regulate floral bud sex differentiation. Conclusion The present study revealed the differences in morphology and phytohormones content between male and hermaphroditic floral buds of ‘Longyanyeshi 1’ during the process of sex differentiation, and identified a subset of candidate genes and miRNAs putatively associated with its sex differentiation. These findings can provide a foundation for molecular regulatory mechanism researching on andromonoecious persimmon.

Probing the floral developmental stages, bisexuality and sex reversions in castor (Ricinus communis L.)

Castor (Ricinus communis L) is an ideal model species for sex mechanism studies in monoecious angiosperms, due to wide variations in sex expression. Sex reversion to monoecy in pistillate lines, along with labile sex expression, negatively influences hybrid seed purity. The study focuses on understanding the mechanisms of unisexual flower development, sex reversions and sex variations in castor, using various genotypes with distinct sex expression pattern. Male and female flowers had 8 and 12 developmental stages respectively, were morphologically similar till stage 4, with an intermediate bisexual state and were intermediate between type 1 and type 2 flowers. Pistil abortion was earlier than stamen inhibition. Sex alterations occurred at floral and inflorescence level. While sex-reversion was unidirectional towards maleness via bisexual stage, at high day temperatures (Tmax > 38 °C), femaleness was restored with subsequent drop in temperatures. Temperature existing for 2–3 weeks during floral meristem development, influences sexuality of the flower. We report for first time that unisexuality is preceded by bisexuality in castor flowers which alters with genotype and temperature, and sex reversions as well as high sexual polymorphisms in castor are due to alterations in floral developmental pathways. Differentially expressed (male-abundant or male-specific) genes Short chain dehydrogenase reductase 2a (SDR) and WUSCHEL are possibly involved in sex determination of castor.

The SAP function in pistil development was proved by two allelic mutations in Chinese cabbage (Brassica rapa L. ssp. pekinensis)

Background Pistil development is a complicated process in plants, and female sterile mutants are ideal material for screening and cloning pistil development-related genes. Using the female sterile mutant (fsm1), BraA04g009730.3C was previously predicted as a candidate mutant gene encoding the STERILE APETALA (SAP) transcriptional regulator. In the current study, a parallel female sterile mutant (fsm2) was derived from EMS mutagenesis of a Chinese cabbage DH line ‘FT’ seeds. Results Both fsm2 and fsm1 mutant phenotypes exhibited pistil abortion and smaller floral organs. Genetic analysis indicated that the phenotype of mutant fsm2 was also controlled by a single recessive nuclear gene. Allelism testing showed that the mutated fsm1 and fsm2 genes were allelic. A single-nucleotide mutation (G-to-A) in the first exon of BraA04g009730.3C caused a missense mutation from GAA (glutamic acid) to GGA (glycine) in mutant fsm2 plants. Both allelic mutations of BraA04g009730.3C in fsm1 and fsm2 conferred the similar pistil abortion phenotype, which verified the SAP function in pistil development. To probe the mechanism of SAP-induced pistil abortion, we compared the mutant fsm1 and wild-type ‘FT’ pistil transcriptomes. Among the 3855 differentially expressed genes obtained, 29 were related to ovule development and 16 were related to organ size. Conclusion Our study clarified the function of BraA04g009730.3C and revealed that it was responsible for ovule development and organ size. These results lay a foundation to elucidate the molecular mechanism of pistil development in Chinese cabbage.

Analyzing Differentially Expressed Genes and Pathways Associated with Pistil Abortion in Japanese Apricot via RNA-Seq

Reproduction is a critical stage in the flower development process, and its failure causes serious problems affecting fruit quality and yield. Pistil abortion is one of the main factors in unsuccessful reproduction and occurs in many fruit plants. In Japanese apricot, the problem of pistil abortion is very common and affects fruit quality and plant yield; however, its molecular mechanism is not clearly understood. Therefore, in the current study, we used RNA-Seq to identify the differentially expressed genes (DEGs) and pathways actively involved in pistil abortion. A total of 3882 differentially expressed genes were found after cutoff and pairwise comparison analysis. According to KEGG pathway analysis, plant hormone signaling transduction and metabolic pathways were found most significantly enriched in this study. A total of 60 transcription factor families such as MADS-box, NAC and TCP showed their role in this process. RT-qPCR assays confirmed that the expression levels were consistent with RNA-Seq results. This study provides an alternative to be considered for further studies and understanding of pistil abortion processes in Japanese apricot, and it provides a reference related to this issue for other deciduous fruit crops.

Microstructural Observation and Transcriptome Analysis of Pistil Abortion in ‘Li Guang’ Apricot

Background: ‘Li Guang’ apricot, a famous local variety, originated in Dunhuang city, Gansu Province,China. It has a long flowering period and a large amount of flowers, but serious pistil abortion has become one of the key factors affecting the fruit set, yield and quality. The distribution and regulation of hormones play an important role in signal molecules of flower abortion. The critical mechanisms of hormone metabolism and the expression levels of genes involved in these processes are, however, poorly understood. Results: To clarify the critical molecular mechanisms of hormone-induced abortion in apricot, normal and abortive flower buds were taken as materials, the pistil abortion of apricot flower was studied by paraffin section, and the RNA seq was used to identify the genes related to flowering regulation. The pistil style was lower than filament. Microstructure showed that the pollen grains of abortive flowers were decreased sharply, the ovaries shrunk and the ovule primordia developed stagnately. Through RNA-Seq, 6647 differentially expressed genes, including 2543 up-regulated and 4104 down-regulated genes, were identified. According to the KEGG Pathway, the pyruvate metabolism, plant hormone signal transduction, spliceosome, RNA transport, protein processing in endoplasmic reticulum and other metabolic pathways were significantly enriched. It revealed that AUX1, AUX / IAA, TIR1, ARF, GH3 and SAUR , vital genes displayed identical differential expression profiles to auxin transduction pathway, and ABF , SnRK2 , PP2C to abscisic acid, JAZ, MYC2 to jasmonic acid. The qRT-PCR assay with independent samples showed that the expression levels of these selected genes were basically consistent with RNA-Seq results. Conclusions : In the whole differentiate stage of flower, pistil abortion represent versatile style . In this process, the changes of hormones play an important role in pistil abortion, especially IAA,GA,and CTK. Related genes involved in hormones synthesis expression regulate the content of hormones and to adapt to the occurrence of pistil abortion under adversity. At the same time, the ethylene response signal factor ERF1/2 (DN70415) was up-regulated in normal flowers, which further indicated that ethylene might be the key regulatory factor affecting the abortion of ‘Liguang’ apricot flowers.