Filtrates from cultures of endophytic fungi isolated from leaves of Copaifera oblongifolia (Fabaceae) affect germination and seedling development differently

Interactions between endophytic fungi (EFs) and their host plants range from positive to neutral to negative. The results of such interactions can vary depending on the organ of the infected host plant. EFs isolated from the leaves of some species of plants have potential for use as agents to inhibit seed germination and control invasive plants. The objectives of this study were to identify EFs present in the leaves of Copaifera oblongifolia and to evaluate the role of these fungi in seed germination and seedling development. A total of 11 species of EFs were isolated, which were identified using the internal transcribed spacers (ITS) sequence of the nuclear ribosomal DNA. The isolated species of EFs are generalists and probably are transmitted horizontally. Laboratory tests revealed that filtrates of these fungal isolates differently affect seed germination and seedling development of C. oblongifolia. The species Curvularia intermedia, Neofusicoccum parvum, Pseudofusicoccum stromaticum and Phomopsis sp. negatively affected seed germination, with N. parvum standing out for its negative effects, inhibiting seedling germination and survival in 89 and 222%, respectively. In addition, Cochliobolus intermedius negatively affected seedling development. Thus, the combined use of N. parvum and C. intermedius, or products from the metabolism of these microorganisms, in the control of invasive plants deserves attention from future studies.

R2R3-MYB Transcription Factor NtMYB330 Regulates Proanthocyanidin Biosynthesis and Seed Germination in Tobacco (Nicotiana tabacum L.)

Proanthocyanidins (PAs) are important phenolic compounds and PA biosynthesis is regulated by a ternary MBW complex consisting of a R2R3-MYB regulator, a bHLH factor and a WDR protein. In this study, a tobacco R2R3-MYB factor NtMYB330 was characterized as the PA-specific regulator in which the PA biosynthesis was promoted in the flowers of NtMYB330-overexpressing lines while decreased in the flowers of ntmyb330 mutants. NtMYB330 can interact with flavonoid-related bHLH partner NtAn1b and WDR protein NtAn11-1, and the NtMYB330-NtAn1b complex is required to achieve strong transcriptional activation of the PA-related structural genes NtDFR1, NtANS1, NtLAR1 and NtANR1. Our data reveal that NtMYB330 regulates PA biosynthesis in seeds and affects seed germination, in which NtMYB330-overexpressing lines showed higher PA accumulations in seed coats and inhibited germination, while ntmyb330 mutants had reduced seed coat PAs and improved germination. NtMYB330 affects seed germination possibly through two mechanisms: modulating seed coat PAs to affect coat-imposed dormancy. In addition, NtMYB330 regulates the expressions of abscisic acid (ABA) and gibberellin acid (GA) signaling-related genes, affecting ABA-GA crosstalk and seed germination. This study reveals that NtMYB330 specifically regulates PA biosynthesis via formation of the MBW complex in tobacco flowers and affects germination through adjustment of PA concentrations and ABA/GA signaling in tobacco seeds.

Impact of Environmental Factors on Seed Germination and Seedling Emergence of White Clover (Trifolium repens L.)

White clover (Trifolium repens L.) is cultivated as a forage crop and planted in various landscapes for soil conservation. There are numerous reports of failed white clover stands each year. A good understanding of the seed germination biology of white clover in relation to environmental factors is essential to achieve successful stand establishment. A series of experiments were conducted to investigate the impacts of light, temperature, planting depth, drought, and salt stress on seed germination and the emergence of white clover. White clover is negatively photoblastic, and seed germination averaged 63 and 66% under light and complete dark conditions 4 weeks after planting (WAP), respectively. Temperature affected the seed germination speed and rate. At 1 WAP, seeds incubated at 15 to 25 °C demonstrated a significantly higher germination rate than the low temperatures at 5 and 10 °C; however, the germination rate did not differ among the temperature treatments at 4 WAP. The results suggest that white clover germination decreases with increasing sowing depths, and the seeds should be sown on the soil surface or shallowly buried at a depth ≤1 cm to achieve an optimal emergence. White clover seeds exhibited high sensitivity to drought and salinity stress. The osmotic potential and NaCl concentration required to inhibit 50% seed germination were −0.19 MPa and 62.4 mM, respectively. Overall, these findings provide quantifiable explanations for inconsistent establishment observed in field conditions. The results obtained in this research can be used to develop effective planting strategies and support the successful establishment of white clover stands.