Chloroplast is a discrete, highly structured, and semi-autonomous cellular organelle. The small genome of chloroplast makes it an up-and-coming platform for synthetic biology. As a special means of synthetic biology, chloroplast genetic engineering shows excellent potential in reconstructing various sophisticated metabolic pathways within the plants for specific purposes, such as improving crop photosynthetic capacity, enhancing plant stress resistance, and synthesizing new drugs and vaccines. However, many plant species exhibit limited efficiency or inability in chloroplast genetic transformation. Hence, new transformation technologies and tools are being constantly developed. In order to further expand and facilitate the application of chloroplast genetic engineering, this review summarizes the new technologies in chloroplast genetic transformation in recent years and discusses the choice of appropriate synthetic biological elements for the construction of efficient chloroplast transformation vectors.
Sucrose phosphate synthase (SPS), as a rate-limiting enzyme working in conjunction with sucrose-6-phosphate phosphatase (SPP) for sucrose synthesis, plays an essential role in energy provision during growth and development as well as improving fruit quality in plants. However, the systematic analysis and evolutionary pattern of the SPS gene family in apple still lacking. In the present study, a total of seven MdSPS and four MdSPP genes were identified from the Malus domestica genome GDDH13 v1.1. The gene structures and their promoter cis-elements, protein conserved motifs, subcellular localizations, physiological and biochemical properties were analyzed. Chromosome location and gene-duplication analysis demonstrated that whole-genome duplication (WGD) and segmental duplication played vital roles in MdSPS gene family expansion. The Ka/Ks ratio of pairwise MdSPS genes indicated that the members of this family have suffered from strong purifying selection during domestication. Furthermore, three SPS gene subfamilies were classified on the basis of phylogenetic relationship, old gene duplication and significantly divergent evolutionary rates among the SPS gene subfamilies were observed. In addition, a major gene MdSPSA2.3 related to sucrose accumulation were identified according to the highly consistent change trends of its expression in four apple varieties (‘Golden Delicious’, ‘Fuji’, ‘Qinguan’ and ‘Honeycrisp’) and the correlation between gene expression and soluble sugar content during fruit development. Further, the virus-induced silencing of MdSPSA2.3 confirmed the function of MdSPSA2.3 in sucrose accumulation of apple fruit. The present study lays a theoretical foundation to better clarify the biological functions of the MdSPS genes during apple fruit development.
In recent years, Meloidogyne enterolobii has emerged as a major parasitic nematode infesting many plants in tropical or subtropical areas. However, the regions of potential distribution and the main contributing environmental variables for this nematode are unclear. Under the current climate scenario, we predicted the potential geographic distributions of M. enterolobii worldwide and in China using a Maximum Entropy (MaxEnt) model with the occurrence data of this species. Furthermore, the potential distributions of M. enterolobii were projected under three future climate scenarios (BCC-CSM2-MR, CanESM5 and CNRM-CM6-1) for the periods 2050s and 2090s. Changes in the potential distribution were also predicted under different climate conditions. The results showed that highly suitable regions for M. enterolobii were concentrated in Africa, South America, Asia, and North America between latitudes 30° S to 30° N. Bio16 (precipitation of the wettest quarter), bio10 (mean temperature of the warmest quarter), and bio11 (mean temperature of the coldest quarter) were the variables contributing most in predicting potential distributions of M. enterolobii. In addition, the potential suitable areas for M. enterolobii will shift toward higher latitudes under future climate scenarios. This study provides a theoretical basis for controlling and managing this nematode.
In the U.S., Helicoverpa zea (Boddie) is a major pest targeted by both transgenic maize and cotton expressing Bacillus thuringiensis (Bt) proteins. Resistance of insect to Bt maize and cotton containing cry1A and cry2A genes has widely occurred in the U.S. In this study, two trials were performed to investigate larval survival and development of a Cry1A.105/Cry2Ab2 dual-protein resistant (VT2P-RR), a susceptible, and an F1 heterozygous (VT2P-RS) populations of H. zea on ears of nine Bt and three non-Bt maize hybrids. The Bt maize hybrids evaluated represent five common pyramided traits expressing two or three of the Cry1A.105, Cry1Ab, Cry1F, Cry2Ab2, and Vip3Aa20 proteins. In the laboratory, neonates of the three H. zea populations were inoculated on silks of ears collected from maize at R1-R2 plant stages; and larval survivorship was checked 10 d after neonate release. All three insect populations survived normally on non-Bt maize ears. Varied numbers of VT2P-RR and VT2P-RS survived on ears of Cry1A.105/Cry2Ab2 maize, while all larvae of the three populations died or could not develop on ears of Vip3Aa20-expressing maize. The results demonstrated that the dual-protein resistant H. zea was not cross-resistant to Vip3Aa20-expressing maize, and thus traits with vip3Aa20 gene should be effective to manage Cry1A.105/Cry2Ab2-resistant H. zea. The resistance in VT2P-RR was determined to be incomplete on Cry1A.105/Cry2Ab2 maize. The effective dominance levels varied greatly, from recessive to incompletely dominant, depending on maize hybrids and trials, suggesting that proper selection of maize hybrids could be important for mitigating the Cry1A.105/Cry2Ab2 resistance. The data generated should aid in modeling multiple-protein Bt resistance in H. zea.
Spodoptera frugiperda (Lepidoptera: Noctuidae) is an important migratory agricultural pest worldwide, which has invaded many countries in the Old World since 2016 and now poses a serious threat to world food security. The present monitoring and early warning strategies for the fall army worm (FAW) mainly focus on adult population density, but lack an information technology platform for precisely forecasting the reproductive dynamics of the adults. In this study, to identify the developmental status of the adults, we first utilized female ovarian images to extract and screen five features combined with the support vector machine (SVM) classifier and employed male testes images to obtain the testis circular features. Then, we established models for the relationship between oviposition dynamics and the developmental time of adult reproductive organs using laboratory tests. The results show that the accuracy of female ovary development stage determination reached 91%. The mean standard error (MSE) between the actual and predicted values of the ovarian developmental time was 0.2431, and the mean error rate between the actual and predicted values of the daily oviposition quantity was 12.38%. The error rate for the recognition of testis diameter was 3.25%, and the predicted and actual values of the testis developmental time in males had an MSE of 0.7734. A WeChat applet for identifying the reproductive developmental state and predicting reproduction of S. frugiperda was developed by integrating the above research results, and it is now available for use by anyone involved in plant protection. This study developed an automated method for accurately forecasting the reproductive dynamics of S. frugiperda populations, which can be helpful for the construction of a population monitoring and early warning system for use by both professional experts and local people at the county level.
