Effects of LED photoperiods and light qualities on in vitro growth and chlorophyll fluorescence of Cunninghamia lanceolata.

BACKGROUND: Cunninghamia lanceolata (C. lanceolata) is the main fast-growing timber species in southern China. As an alternative to conventional lighting systems, LED has been demonstrated to be an artificial flexible lighting source for commercial micropropagation. The application of LED can provide rapid propagation of C. lanceolata in vitro culture. RESULTS: We applied two-factor randomized block design to study the effects of LED photoperiods and light qualities on the growth and chlorophyll fluorescence of C. lanceolata in vitro culture plantlets. In this study, plantlets were exposed to 20 µmol·m- 2·s- 1 irradiance for three photoperiods, 8, 16, and 24 h under the three composite lights, 88.9% red+ 11.1% blue (R/B), 80.0% red+ 10.0% blue+ 10.0% purple (R/B/P), 72.7% red+ 9.1% blue+ 9.1% purple+ 9.1% green (R/B/P/G), as well as white light (12.7% red+ 3.9% blue+ 83.4% green, W) as control. The results showed that: (1) Plant height, dry weight, rooting rate, average root number, length, surface area and volume, chlorophyll, and chlorophyll fluorescence parameters were significantly affected by photoperiods, light qualities and their interactions. (2) Plantlets subjected to photoperiod 16 h had longer root, higher height, rooting rate, root number, and the higher levels of chlorophyll, chlorophyll a/b, Y (II), qP, NPQ/4 and ETRII compared to photoperiods 8 h and 24 h, while Fv/Fm during photoperiod 16 h was lower than 8 h and 24 h. Plantlets exposed to R/B/P/G generated more root and presented higher chlorophyll, Fv/Fo, Y (II), qP, and ETRII than W during photoperiods 8 and 16 h. (3) Total chlorophyll content and ETRII were significant correlated with rooting rate, root length and root volume, while Fv/Fm and ETRII were significant correlated with plant height, average root number and root surface area. (4) 16-R/B/P/G is best for growing C. lanceolata plantlets in vitro. CONCLUSIONS: This study demonstrated the effectiveness of photoperiods and light qualities using LEDs for micropropagation of C. lanceolata. The best plantlets were harvested under 16-R/B/P/G treatment. And there was a correlation between the growth and the chlorophyll and chlorophyll fluorescence of their leaves under different photoperiod and light quality. These results can contribute to improve the micropropagation process of this species.

Effects of Composite LED Light on Root Growth and Antioxidant Capacity of Cunninghamia lanceolata Tissue Culture Seedlings.

We used the 12th generation of the Cunninghamia (C.) lanceolata tissue culture seedlings, and white light emitting diode (LED) light as control (CK). We applied five composite LED light treatments, red-blue 4:1, 8:1 (4R1B and 8R1B), red-blue-purple 8:1:1 (8R1B1P), and red-blue-purple-green 6:1:1:1, 8:1:1:1 (6R1B1P1G and 8R1B1P1G), to study the effects of light quality on root growth characteristics and antioxidant capacity of C. lanceolata tissue culture seedlings. The results showed that: (1) rooting rate, average root number, root length, root surface area, and root activity were higher with 6R1B1P1G and 8R1B1P1G treatments compared to 4R1B, 8R1B, 8R1B1P and CK treatments; and the root growth parameters under the 8R1B1P1G treatment were as high as 95.50% for rooting rate, 4.63 per plant of the average number of root, 5.95 cm root length, 1.92 cm2 surface area, and 145.56 ng/(g·h) root activity, respectively. (2) The composite lights of 4R1B, 8R1B, 8R1B1P, 6R1B1P1G, and 8R1B1P1G are beneficial for the accumulation of soluble sugar content (SSC) and soluble protein content (SPC), but not conducive for the increase of free proline content (FPC); the plants under 6R1B1P1G and 8R1B1P1G treatments had higher superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX) activity and lower malondialdehyde (MDA), polyphenol oxidase (PPO) activity. (3) Redundancy analysis showed that POD activity positively correlated with root activity; SPC, SOD and CAT activities positively correlated with root growth parameters; while SSC, MDA content, APX and PPO activities negatively correlated with root growth parameters. These results suggest that the responses of root growth and antioxidant capacity of the C. lanceolata tissue culture seedlings to different light qualities vary. The relationship between root growth parameters and antioxidant capacity was closely related. Red-blue-purple-green was the most suitable composite light quality for root growth of C. lanceolata tissue culture seedlings, and 8:1:1:1 was the optimal ratio, under which the rooting rate, root activity and root growth of tissue culture seedlings peaked.

3D reconstruction and capsid protein characterization of grass carp reovirus.

Grass carp reovirus (GCRV) is a relatively new virus first isolated in China and is a member of the Aquareovirus genus of the Reoviridae family. Recent report of genomic sequencing showed that GCRV shared high degree of homology with mammalian reovirus (MRV). As a step of our effort to understand the structural basis of GCRV pathogenesis, we determined the three-dimensional (3D) structure of GCRV capsid at 17 A resolution by electron cryomicroscopy. Each GCRV capsid has a multilayered organization, consisting of an RNA core, an inner, middle and outer protein layer. The outer layer is made up of 200 trimers that are arranged on an incomplete T=13 icosahedral lattice. A characteristic feature of this layer is the depression resulting from the absence of trimers around the peripentonal positions, revealing the underlying trimers on the middle layer. There are 120 subunits in the inner layer arranged with T=1 symmetry. These structural features are common to other members of the Reoviridae. Moreover, SDS-PAGE analysis showed that GCRV virions contain seven structural proteins (VP1-VP7). These structural proteins have a high degree of sequence homology to MRV, consistent with the structural similarities observed in our study. The high structural similarities of isolated GCRV and MRV suggest that future structural studies focusing on GCRV entering into and replicating within its host cell are necessary in order to fully understand the structural basis of GCRV pathogenesis.

Web portal to an image database for high-resolution three-dimensional reconstruction.

The exponential increase of image data in high-resolution reconstructions by electron cryomicroscopy (cryoEM) has posed a need for efficient data management solutions in addition to powerful data processing procedures. Although relational databases and web portals are commonly used to manage sequences and structures in biological research, their application in cryoEM has been limited due to the complexity in accomplishing the dual tasks of interacting with proprietary software and simultaneously providing data access to users without database knowledge. Here, we report our results in developing web portal to SQL image databases used by the Image Management and Icosahedral Reconstruction System (IMIRS) to manage cryoEM images for subnanometer-resolution reconstructions. Fundamental issues related to the design and deployment of web portals to image databases are described. A web browser-based user interface was designed to accomplish data reporting and other database-related services, including user authentication, data entry, graph-based data mining, and various query and reporting tasks with interactive image manipulation capabilities. With an integrated web portal, IMIRS represents the first cryoEM application that incorporates both web-based data reporting tools and a complete set of data processing modules. Our examples should thus provide general guidelines applicable to other cryoEM technology development efforts.

IMIRS: a high-resolution 3D reconstruction package integrated with a relational image database.

Recent advances in electron cryomicroscopy instrumentation and single particle reconstruction have created opportunities for high-throughput and high-resolution three-dimensional (3D) structure determination of macromolecular complexes. However, it has become impractical and inefficient to rely on conventional text file data management and command-line programs to organize and process the increasing numbers of image data required in high-resolution studies. Here, we present a distributed relational database for managing complex datasets and its integration into our high-resolution software package IMIRS (Image Management and Icosahedral Reconstruction System). IMIRS consists of a complete set of modular programs for icosahedral reconstruction organized under a graphical user interface and provides options for user-friendly, step-by-step data processing as well as automatic reconstruction. We show that the integration of data management with processing in IMIRS automates the tedious tasks of data management, enables data coherence, and facilitates information sharing in a distributed computer and user environment without significantly increasing the time of program execution. We demonstrate the applicability of IMIRS in icosahedral reconstruction toward high resolution by using it to obtain an 8-A 3D structure of an intermediate-sized dsRNA virus.