Develop a preliminary core germplasm with the novel polymorphism EST-SSRs derived from three transcriptomes of colored calla lily (Zantedeschia hybrida).

The development of high-throughput sequencing technology has made it possible to develop molecular markers such as EST-SSR from transcriptome sequences in non-model plants such as bulbous flowers. However, the EST-SSR markers that have been developed are weakly validated and low polymorphic due to the short read size and poor quality of the assembled sequences. This study therefore used the CandiSSR pipeline to identify 550 potential polymorphic SSR loci among 487 homologous unigenes based on the transcriptomic sequences of three varieties of colored calla lily, and 460 of these loci with appropriate flanking sequences were suitable for primer pairs design. A further validation with 200 randomly selected EST-SSRs demonstrated an increase of more than 30% and 100% in amplification validity and polymorphism, respectively, in comparison with our previous study. In addition, since most of the current varieties of colored calla lily are hybridized from a few species, which have low genetic diversity, we subsequently identified primary core germplasm for 160 colored calla lily accessions using the aforementioned 40 polymorphic EST-SSRs. It was concluded that the core germplasm containing 42 accessions derived from the M strategy incorporated into the software Power Core was the most representative of all 160 original germplasm, as evidenced by the preservation of 100% of the EST-SSR variation, with a higher level of genetic diversity and heterogeneity (Nei = 0.40, I = 0.66, PIC = 0.43). This study provides a practical example of polymorphism EST-SSR markers developed from multiple transcriptomes for non-model plants. A future breeding program for colored calla lily will also benefit from the core germplasm defined by those molecular markers.

Establishment of an efficient regeneration and Agrobacterium transformation system in mature embryos of calla lily (Zantedeschia spp.).

Calla lily (Zantedeschia spp.) have great aesthetic value due to their spathe-like appearance and richness of coloration. However, embryonic callus regeneration is absent from its current regeneration mechanism. As a result, constructing an adequate and stable genetic transformation system is hampered, severely hindering breeding efforts. In this research, the callus induction effectiveness of calla lily seed embryos of various maturities was evaluated. The findings indicated that mature seed embryos were more suitable for in vitro regeneration. Using orthogonal design experiments, the primary elements influencing in vitro regeneration, such as plant growth regulators, genotypes, and nanoscale materials, which was emergent uses for in vitro regeneration, were investigated. The findings indicated that MS supplemented with 6-BA 2 mg/L and NAA 0.1 mg/L was the optimal medium for callus induction (CIM); the germination medium (GM) was MS supplemented with 6-BA 2 mg/L NAA 0.2 mg/L and 1 mg/L CNTs, and the rooting medium (RM) was MS supplemented with 6-BA 2 mg/L NAA 0.7 mg/L and 2 mg/L CNTs. This allowed us to verify, in principle, that the Agrobacterium tumefaciens-mediated genetic transformation system operates under optimal circumstances using the GUS reporter gene. Here, we developed a seed embryo-based genetic transformation regeneration system, which set the stage for future attempts to create new calla lily varieties.

[Dry and Wet Deposition of Atmospheric Nitrogen in Small Catchments].

Atmospheric nitrogen deposition is one of the important pathways for the transmission of terrestrial pollutants and nutrients to aquatic ecosystems. A considerable amount of nitrogen substances is delivered into the surface water environment via atmospheric deposition in the catchments, which are greatly affected by human activities, and have serious negative effects on the nutrient level and aquatic habitat environment. The Shixia Catchment north of the Miyun Reservoir was selected as study area. Samples of particulate dry and wet deposition were collected and tested. Subsequently, the variation of the atmospheric nitrogen deposition flux and factors impacting it were determined; ultimately, the contribution of atmospheric nitrogen deposition to the nitrogen budget of the catchment was estimated. The major results are as follows:① The total nitrogen deposition flux of particulate dry and wet deposition is characterized by a significant temporary variation. Regarding the total nitrogen, the ammonia nitrogen and wet deposition flux are the highest in summer and dissolved organic nitrogen is the highest in spring. However, the flux of nitrate nitrogen shows no notable seasonal fluctuation. The general trend of the particulate dry deposition flux is supreme in winter, followed by that in autumn and spring. In detail, the total nitrogen and ammonia nitrogen deposition are the largest in winter, while the seasonal variance of nitrate nitrogen appears to be less apparent. The dry deposition flux of dissolved organic nitrogen is the greatest in autumn. ② The total flux of nitrogen deposition is 43.14 kg·hm-2 in the Shixia Catchment; the wet and dry deposition flux account for 39.85% and 60.15%, respectively. ③ Rainfall and wind speed conditions are important factors impacting the atmospheric nitrogen deposition. The rainfall and rainfall intensity are negatively correlated with the wet nitrogen deposition concentration. With respect to the particulate dry deposition, the average wind speed during the monitoring period is important to ammonia nitrogen. ④ The contribution of atmospheric nitrogen deposition to the total nitrogen input in this catchment is approximately 15.09%, second only to livestock and poultry farming and rural life. The results of this study provide information that is useful for the nitrogen management in the catchment.