Histological and Cytological Characterization of Anther and Appendage Development in Asian Lotus (Nelumbo nucifera Gaertn.).

The lotus (Nelumbo Adans.) is a perennial aquatic plant with important value in horticulture, medicine, food, religion, and culture. It is rich in germplasm and more than 2000 cultivars have been cultivated through hybridization and natural selection. Microsporogenesis and male gametogenesis in the anther are important for hybridization in flowering plants. However, little is known about the cytological events, especially related to the stamen, during the reproduction of the lotus. To better understand the mechanism controlling the male reproductive development of the lotus, we investigated the flower structure of the Asian lotus (N. nucifera). The cytological analysis of anther morphogenesis showed both the common and specialized cytological events as well as the formation of mature pollen grains via meiosis and mitosis during lotus anther development. Intriguingly, an anatomical difference in anther appendage structures was observed between the Asian lotus and the American lotus (N. lutea). To facilitate future study on lotus male reproduction, we categorized pollen development into 11 stages according to the characterized cytological events. This discovery expands our knowledge on the pollen and appendage development of the lotus as well as improving the understanding of the species differentiation of N. nucifera and N. lutea.

Chromosome Nomenclature and Cytological Characterization of Sacred Lotus.

Sacred lotus is a basal eudicot plant that has been cultivated in Asia for over 7,000 years for its agricultural, ornamental, religious, and medicinal importance. A notable characteristic of lotus is the seed longevity. Extensive endeavors have been devoted to dissect its genome assembly, including the variety China Antique, which germinated from a 1,300-year-old seed. Here, cytogenetic markers representing the 10 largest megascaffolds, which constitute approximately 70% of the lotus genome assembly, were developed. These 10 megascaffolds were then anchored to the corresponding lotus chromosomes by fluorescence in situ hybridization using these cytogenetic markers, and a set of chromosome-specific cytogenetic markers that could unambiguously identify each of the 8 chromosomes was generated. Karyotyping was conducted, and a nomenclature based on chromosomal length was established for the 8 chromosomes of China Antique. Comparative karyotyping revealed relatively conserved chromosomal structures between China Antique and 3 modern cultivars. Interestingly, significant variations in the copy number of 45S rDNA were detected between China Antique and modern cultivars. Our results provide a comprehensive view on the chromosomal structure of sacred lotus and will facilitate further studies and the genome assembly of lotus.

Induction and quantitative proteomic analysis of cell dedifferentiation during callus formation of lotus (Nelumbo nucifera Gaertn.spp. baijianlian).

Lotus is an aquatic plant with high nutritional, ornamental and medical values. Its callus formation is crucial for germplasm innovation by genetic transformation. In this study, embryogenic callus was successfully induced on appropriate medium using cotyledons at 12days after pollination as explants. To dissect cellular dedifferentiation and callus formation processes at the proteome level, cotyledons before and tissues from 10 to 20days after induction were sampled for shotgun proteomic analysis. By applying multivariate statistics 91 proteins were detected as differentially regulated, and sorted into 6 functional groups according to MapMan ontology analysis. Most of these proteins were implicated in various metabolisms, demonstrating that plant cells underwent metabolism reprogramming during callus induction. 14.3% proteins were associated with stress and redox, indicating that the detached explants were subjected to a variety of stresses; 13.2% were cell and cell wall-related proteins, suggesting that these proteins played important roles in rapid cell division and proliferation. Some proteins were further evaluated at the mRNA levels by quantitative reverse transcription PCR analysis. In conclusion, the results contributed to further deciphering of molecular processes of cellular dedifferentiation and callus formation, and provided a reference data set for the establishment of transgenic transformation in lotus.

Fabrication of left-handed metal microcoil from spiral vessel of vascular plant.

Silver microcoil is fabricated through a biotemplating process combined with electroless plating. Spiral vessels in Lotus root are employed as a biotemplate because of their left-handed coil structure. The silver microcoil exhibits a solenoidal microcoil showing self-inductance in the level of picohenry, which could be applied for electromagnetic-responsive materials in the high-frequency region such as millimeter waves or terahertz waves.

Changes in extreme high-temperature tolerance and activities of antioxidant enzymes of sacred lotus seeds.

Sacred lotus (Nelumbo nucifera Gaertn. 'Tielian') seed is long-lived and extremely tolerant of high temperature. Water content of lotus and maize seeds was 0.103 and 0.129 g H2O [g DW](-1), respectively. Water content, germination percentage and fresh weight of seedlings produced by surviving seeds gradually decreased with increasing treatment time at 100 degrees C. Germination percentage of maize (Zea mays L. 'Huangbaogu') seeds was zero after they were treated at 100 degrees C for 15 min and that of lotus seeds was 13.5% following the treatment at 100 degrees C for 24 h. The time in which 50% of lotus and maize seeds were killed by 100 degrees C was about 14.5 h and 6 min, respectively. With increasing treatment time at 100 degrees C, relative electrolyte leakage of lotus axes increased significantly, and total chlorophyll content of lotus axes markedly decreased. When treatment time at 100 degrees C was less than 12 h, subcellular structure of lotus hypocotyls remained fully intact. When treatment time at 100 degrees C was more than 12 h, plasmolysis gradually occurred, endoplasmic reticulum became unclear, nuclei and nucleoli broke down, most of mitochondria swelled, lipid granules accumulated at the cell periphery, and organelles and plasmolemma collapsed. Malondialdehyde (MDA) content of lotus axes and cotyledons decreased during 0 -12 h of the treatment at 100 degrees C and then increased. By contrast, the MDA content of maize embryos and endosperms increased during 5-10 min of the treatment at 100 degrees C and then decreased slightly. For lotus seeds: (1) activities of superoxide dismutase (SOD) and glutathione reductase (GR) of axes and cotyledons and of catalase (CAT) of axes increased during the early phase of treatment at 100 degrees C and then decreased; and (2) activities of ascorbate peroxidase (APX) and dehydroascorbate reductase (DHAR) of axes and cotyledons and of CAT of cotyledons gradually decreased with increasing treatment time at 100 degrees C. For maize seeds: (1) activities of SOD and DHAR of embryos and endosperms and of GR of embryos increased during the early phase of the treatment at 100 degrees C and then decreased; and (2) activities of APX and CAT of embryos and endosperms and of GR of endosperms rapidly decreased with increasing treatment time at 100 degrees C. With decrease in seed germination, activities of SOD, APX, CAT, GR and DHAR of axes and cotyledons of lotus seeds decreased slowly, and those of embryos and endosperms of maize seeds decreased rapidly.