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.

Wheat stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is a potentially devastating fungal disease of wheat worldwide.  The present study was to evaluate the resistance of 42 wheat monogenic lines with known stem rust resistance (Sr) genes and 69 wheat cultivars to three new Pgt races (34C0MRGQM, 34C3MKGQM, and 34C6MTGSM) identified from aeciospores at the seedling and adult-plant stages.  The phenotyping results revealed that monogenic lines harboring resistance genes Sr9e, Sr17, Sr21, Sr22, Sr26, Sr30, Sr31, Sr33, Sr35, Sr36, Sr37, Sr38, Sr47, SrTmp, and SrTt3 were effectively resistant to all three Pgt races at the seedling and adult-plant stages.  In contrast, monogenic lines containing Sr5, Sr6, Sr7b, Sr9a, Sr9d, Sr9f, Sr9g, Sr9b, Sr16, Sr24, Sr28, and Sr39 were highly susceptible to these races at both seedling and adult-plant stages.  The other lines with Sr8a, Sr10, Sr11, Sr13, Sr14, Sr15, Sr18, Sr20, Sr19, Sr23, Sr25, Sr27, Sr29, Sr32, and Sr34, displayed variable levels of resistance to one or two of the tested races.  Seedling infection types (ITs) and adult-plant infection responses (IRs) indicated that 41 (59.4%) of the wheat cultivars showed high resistance to all the three races.  Molecular marker analysis showed that four wheat culitvars likely carried Sr2, 20 wheat culitvars likely carried Sr31, 9 wheat culitvars likely carried Sr38, and none of the cultivars carried Sr24, Sr25, and Sr26.  Our results provide a scientific basis for rational utilization of the tested Sr genes and wheat cultivars against these novel Pgt races. 

The efficacies of biological and conventional chemical insecticides against two major insect pests of alfalfa (aphids and thrips) were compared in three sites across China’s alfalfa belt.  In addition, the persistence of the residues of chemical insecticides in alfalfa and their influence on the quality of alfalfa hay were examined.  Efficacy varied among the different biological and chemical insecticides.  The chemical insecticides were significantly more effective than biopesticides in a short time-frame.  The efficacy period of biopesticides was significantly longer than that of chemical insecticides, and the corrected mortality rate of aphids in all regions was above 50% at 14 days after application.  The analysis of pesticide residues showed that the residual doses of all the pesticides were within the allowed ranges after the safe period.  The acid detergent fiber and neutral detergent fiber contents in alfalfa hay were higher and the protein content was lower in chemical insecticide treatments than in biopesticide treatments in Hebei.  The relative feeding value of alfalfa hay treated with Metarhizium anisopliae IPP330189 was the highest among the treatments.  Compared with chemical insecticides, the yield of alfalfa hay was higher in the biopesticides treatments.  Biopesticides show a stronger control effect on insect populations and also a better improvement in the quality of alfalfa hay than chemical insecticides.  This study provides a basis for exploring and developing a comprehensive control regime for alfalfa insect pests in the different alfalfa-growing regions in China, and for reducing chemical insecticide usage and improving forage quality.

The sugar beet cyst nematode, Heterodera schachtii, is a major parasite of sugar beet which has been recognized and listed as a quarantine nematode in China and more than 20 countries and regions worldwide.  A survey for important nematodes was undertaken in the sugar beet planting area of China during 2015–2018, and numerous cysts were collected from sugar beet fields in Xinyuan County, Xinjiang Uygur Autonomous Region of China.  The observations of morphological and morphometric characteristics revealed that cysts, vulval cones and second-stage juveniles of the Xinjiang population were in the same range of each other and within those of other reported H. schachtii populations.  Molecular analysis of rDNA-ITS, 28S-D2/D3 and mtDNA cytochrome c oxidase subunit 1 (COI) gene sequences suggested that the Xinjiang population clustered in a branch with those foreign populations, and the sequence similarity was as high as 99.81–100%.  Moreover, this result was confirmed by PCR assay with species-specific primer SHF6 and rDNA2 of H. schachtii, and the pathogenicity test confirmed successful Xinjiang population reproduction in both plant hosts.  In conclusion, based on morphological and molecular characterization, this study confirmed that the cyst nematode population collected from sugar beet fields in Xinjiang is H. schachtii.  As far as we know, this is the first report of H. schachtii on sugar beets in Xinjiang, China.

Bread wheat (Triticum aestivum L.) is one of the most important staple crops worldwide.  The phytohormone auxin plays critical roles in the regulation of plant growth and development.  However, only a few auxin-related genes have been genetically demonstrated to be involved in the control of plant architecture in wheat thus far.  In this study, we characterized an auxin-related gene in wheat, TaIAA15, and found that its ectopic expression in rice decreased the plant height and increased the leaf angle.  Correlation analysis indicated that TaIAA15-3B was associated with plant height (Ph), spike length (SL) and 1 000-grain weight (TGW) in wheat, and Hap-II of TaIAA15-3B was the most favored allele and selected by modern breeding in China.  This study sheds light on the role of auxin signaling on wheat plant architecture as well as yield related traits.

Ovine bones are the major by-products after slaughtered.  The present study was conducted to extract and characterize acid soluble collagens (ASC) and pepsin soluble collagens (PSC) from ovine bones (Ujumuqin sheep).  Ovine bones collagen were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) as type I collagen.  The results of Fourier transform infrared (FTIR) spectra analysis testified the existence of triple superhelical structure in both ASC and PSC, showing pepsin did not disrupt the triple helical structure of ovine bones collagen.  Glycine, accounting for one-third of total amino acids, was the major amino acid for ovine bones collagen.  Higher imino acid content was responsible for higher thermal denaturation temperature of ovine bones collagen compared to fish collagens.  The isoelectric point of ASC was lower than PSC due to the higher content of acidic amino acids.  Therefore, this study provides the potential reference for collagen extraction and application of ovine bones by-procduct.

Effect of salicylic acid (SA) on chilling injury (CI) of sponge gourd during storage (9 days, 9°C) plus shelf life (2 days, 20°C) was evaluated in this study.  SA treatment at the concentration of 1.5 mmol L^–1 significantly reduced postharvest CI of sponge gourds.  Besides, the application of SA could effectively decrease the electrolyte leakage, reduce the accumulation of malondialdehyde (MDA) and total phenolics, enhance the activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX), and inhibit the activities of phenylalanine ammonia-lyase (PAL) and polyphenol oxidase (PPO).  The beneficial effects of SA could be attributed to preserved membrane integrity, inhibited membrane peroxidation, enhanced antioxidant system and suppressed activities of browning related enzymes.  In a sense, SA as a postharvest tool may be commercially used in alleviating CI of sponge gourd.

  Food waste results in nutritional losses, ecological damage, and environmental pollution. This survey is conducted in Beijing and aims to determine whether food waste can be reduced by food packing when leftovers are produced at the dining table and to identify factors that affect food packing behavior and the use of leftovers. Based on statistical and econometric analyses of the relationship between leftovers packing and possible factors, this study finds that the age, educational level, employment status, farming experience, environmental protection consciousness, food saving advertisement, families with old members, and reasons for dining out significantly influence the food packing behavior of the consumers. Moreover, the dining environment plays an important role in leftovers packing. People with intimate relationships, such as families, friends, classmates, or colleagues, are more willing to take leftovers home. Business partners do the opposite. Finally, almost all packed leftovers (91.59%) are eaten by people and animals. Therefore, packing leftovers is an excellent approach to reduce food waste.

Blue and red lights differently regulate leaf photosynthesis. Previous studies indicated that plants under blue light generally exhibit better photosynthetic characteristics than those under red light. However, the regulation mechanism of related photosynthesis characteristics remains largely unclear. Here, four light qualities treatments (300 μmol m–2 s–1) including white fluorescent light (FL), blue monochromatic light (B, 440 nm), red monochromatic light (R, 660 nm), and a combination of red and blue light (RB, R:B=8:1) were carried out to investigate their effects on the activity of photosystem II (PSII) and photosystem I (PSI), and photosynthetic electron transport capacity in the leaves of cucumber (Cucumis sativus L.) seedlings. The results showed that compared to the FL treatment, the R treatment significantly limited electron transport rate in PSII (ETRII) and in PSI (ETRI) by 79.4 and 66.3%, respectively, increased non-light induced non-photochemical quenching in PSII (ΦNO) and limitation of donor side in PSI (ΦND) and reduced most JIP-test parameters, suggesting that the R treatment induced suboptimal activity of photosystems and inhibited electron transport from PSII donor side up to PSI. However, these suppressions were effectively alleviated by blue light addition (RB). Compared with the R treatment, the RB treatment significantly increased ETRII and ETRI by 176.9 and 127.0%, respectively, promoted photosystems activity and enhanced linear electron transport by elevating electron transport from QA to PSI. The B treatment plants exhibited normal photosystems activity and photosynthetic electron transport capacity similar to that of the FL treatment. It was concluded that blue light is more essential than red light for normal photosynthesis by mediating photosystems activity and photosynthetic electron transport capacity.

The antifungal activity of chitosan on a common fungal phytopathogen, Sclerotinia sclerotiorum, and the control effect on sclerotinia rot of carrot were investigated. Mycelial growth and fungal biomass were strongly inhibited by chitosan. Using propidium iodide stain combined with fluorescent microscopy, the plasma membrane of chitosan-treated S. sclerotiorum mycelia was observed to be markedly damaged. Concomitantly, protein leakage and lipid peroxidation was also found to be significantly higher in chitosan-treated mycelia compared to the control. Chitosan provided an effective control of sclerotinia rot of carrot, with induction of activity of defense-related enzymes including polyphenoloxidase and peroxidase. These data suggest that the effects of chitosan on sclerotinia rot of carrot may be associated with the direct damage to the plasma membrane and lipid peroxidation of S. sclerotiorum, and the elicitation of defense response in carrot.

Infectious bursal disease virus (IBDV) is responsible for the highly contagious infectious bursal disease of chickens. Further understanding the gene-function is necessary to design the tailored vaccine. The amino acid residue 279, located on strand PF of VP2, is one of the three residues that have been reported to be involved in cell-tropism but with some inconsistency. In this study, to further clarify the amino acids involved in the cell tropism of IBDV, a series of mutations about residue 279 were introduced into the VP2 of vvIBDV Gx strain. With the reverse genetic system, we found single mutation of D279N, double mutations of D279N/A284T or Q253H/D279N were not enough to adapt IBDV to chicken embryo fibroblast (CEF) cell. To evaluate whether residue 279 could influence the replication and virulence of IBDV, the virus rGxHT-279 with three mutations (Q253H/D279N/A284T) was rescued and evaluated. Results showed that the mutation of residue 279 in VP2 had no efficient effects on both the replication efficiency in vitro and the virulence to SPF chickens of IBDV. In summary, the results demonstrated that residue 279 of VP2 did not contribute efficiently to cell tropism, replication efficiency, and virulence of IBDV at least in some strains. These findings provided further information for understanding the gene function of IBDV.

Two experiments were conducted to determine the effects of fiber level from alfalfa meal and sampling time on the apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of amino acids (AA) in growing pigs. A total of 24 ileal-cannulated pigs (Duroc×(Large White×Landrace) with body weight (21.4±1.5) kg) were randomly allotted to 4 treatments. The pigs were provided a corn-soybean meal diet or a diet containing 5, 10 or 20% of alfalfa meal during two 10-d experimental periods. The AID of AA was measured. Six ileal-cannulated pigs were fed a protein-free diet in order to estimate the endogenous protein losses and SID of AA. Ileal AA digestibility was not affected by inclusion of 5 or 10% alfalfa meal in the diet (P>0.05). The AID of His, Lys, Met, Phe, Thr, Val, Ala, Asp, Cys, Gly, Pro, Ser and Tyr reduced by 2.0–6.8% with the addition level of alfalfa meal (linear, P<0.05). The SID of His, Lys, Ser, Thr and Tyr decreased by 2.2–4.3% as the level of alfalfa meal in the diet increased (linear, P<0.05). The AID and SID of AA were not affected by the sampling time (P>0.05). A multiple linear regression analysis, taking into account both the soluble and insoluble fiber content in the diets, explained more than 36% of variation in SID of Ser and Thr (P<0.05). In conclusion, the corn-soybean meal diet containing 10% of alfalfa meal did not affect ileal AA digestion. The AID and SID of AA were similar between two sampling times. Increasing the concentration of total dietary fiber from 12.3 to 21.4% by adding graded levels of alfalfa meal (0–20%) to a corn-soybean meal control diet induced a linear reduction in AID and SID of most AA. Soluble and insoluble fibers from alfalfa meal have differential roles in the AA digestion, which may help explain the variation observed in the SID of partial AA. These findings would provide important information for dietary fiber level and composition related to AA digestion.

Wheat leaf rust (caused by Puccinia triticina) is one of the most important fungal diseases in China. There are tens of winter wheat cultivars which are approved to be released by the government at a national level and more than 100 wheat cultivars at the provincial level. But there is no information about leaf rust (Lr) genes in these cultivars, which makes it difficult for farmers and breeders to select which cultivars they should plant in their fields and use in their breeding programs. The objective of this paper was to identify the leaf rust resistant genes at seedling stage present in the 84 commercial wheat cultivars from China that have been released in the past few years. A set of 20 near isogenic lines with Thatcher background and 6 lines with known Lr genes were used to test the virulence of 12 races of P. triticina (Pt). By comparing the infection types (ITs) produced on the 84 cultivars by the 12 Pt races with the ITs on the differential sets, the Lr genes were postulated. In addition, 8 molecular markers of Lr genes such as Lr9, Lr10, Lr19, Lr20, Lr21, Lr24, Lr26 and Lr29, which are closely linked to or co-segregated with the Lr gene, were used for further validation of the genes in the 84 Chinese winter wheat cultivars. Twelve Lr genes, including Lr1, Lr3, (Lr3bg), (Lr3ka), Lr11, Lr13, Lr14a, Lr16, Lr26, Lr27, Lr30 and Lr31 were postulated to be present either singly or in combinations in these Chinese wheat cultivars. Lr3 and Lr26 were detected most often in the tested cultivars, with frequencies of 51.2 and 38.1%, respectively. No wheat Lr genes were detected in 16 cultivars, and 4 cultivars may carry unknown Lr genes other than those used in this study. Lr9, Lr20, Lr21, Lr24, Lr25 and Lr29 were not present in any of the 84 tested accessions.

The fiber level and composition have an important effect on nutrient digestibility of swine diets. Little information is known about the effects of fiber level and composition from alfalfa meal on nutrient digestibility of fattening pigs fed a corn-soybean meal-based diet. The objective of this experiment was to determine the effects of alfalfa fiber on the growth performance, intestinal nutrient flow and apparent total tract digestibility (ATTD) of nutrients in fattening pigs. 24 barrows (Duroc×(Large White×Landrace), body weight=(60.6±0.7) kg) were randomly allotted to 4 treatments with 6 replicates of 1 pig per replicate. The pigs were provided a control diet or a diet containing 5, 10 or 20% of alfalfa meal during a 14-d experiment period. Average daily gain (ADG) and the ATTD of dry matter (DM), organic matter (OM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF) and gross energy (GE) reduced linearly as the level of alfalfa meal in the diet increased (P<0.01). The total tract flow of DM, OM, CP, NDF, ADF, and GE increased with the increase in dietary alfalfa (linear, P<0.05). Growth performance and nutrient digestion were not affected by inclusion of 5% alfalfa meal in the diet (P>0.05). A multiple linear regression analysis, taking into account both soluble and insoluble fiber intake, explained approximately 70% of the variation in the ATTD of DM, OM, NDF, and GE (P<0.01). In conclusion, alfalfa meal should be limited to less than 5% of the diet in fattening pigs to maximize growth performance and nutrient digestion. Soluble and insoluble fiber from alfalfa meal has the differential roles in nutrient digestion, which may help explain the main variation observed in nutrient digestibility. These findings suggest that knowledge of specific fiber components is necessary to accurately predict the effects of dietary fiber on nutrient digestibility.

Heading date was an important trait that decided the adaptation of wheat to environments. It was modified by genes involved in vernalization response, photoperiod response and development rate. In this study, four loci Xgwm261, Xgwm219, Xbarc23 and Ppd-D1 which were previously reported related to heading time were analyzed based on three groups of wheat including landraces (L), varieties bred before 1983 (B82) and after 1983 (A83) collected from Chinese wheat growing areas. Generally, heading date of landrace was longer than that of varieties. Significant differences in the heading time existed within the groups, which implied that diversification selection was much helpful for adaptation in each wheat zone. Photoperiod insensitive allele Ppd-D1a was the first choice for both landrace and modern varieties, which promoted the heading date about four days earlier than that of sensitive allele Ppd-D1b. The three SSR loci had different characters in the three groups. Predominant allele combination for each zone was predicted for wheat group L and A83, which made great contribution to advantageous traits. Xgwm219 was found to be significantly associated with heading date in Yellow and Huai River Winter Wheat Zone (Zone II) and spike length in Middle and lower Yangtze Valley Winter Wheat Zone (Zone III), which implied functional diversification for adaption. Variation for earliness genes provided here will be helpful for whet breeding in future climatic change.

Stem or black rust of wheat, caused by the fungus Puccinia graminis Pers. f. sp. tritici Eriks. & E. Henn. (Pgt), has historically caused severe losses to wheat (Triticum aestivum L.) production worldwide. In the Fujian and Guangdong provinces of China, six moderate-to-severe epidemics of wheat stem rust have occurred, which caused destructive losses of wheat between 1949 and 1966, although these were brought under control by integrated management. A rapid and reliable detection of the pathogen will contribute to the accurate forecast and seasonal control of this disease. The objective of this study was to develop a diagnostic molecular marker generated from simple sequence repeats (SSR) for the early rapid identification of P. graminis. The genomic DNA of P. graminis, Puccinia striiformis, Puccinia triticina and seven other species was amplified by a pair of SSR primers generated by the FIASCO (fast isolation by AFLP sequences containing repeats) enrichment protocol. The primer set Pgtw (f)/ Pgtw (r) generated a polymorphic pattern displaying a 330-bp DNA fragment specific for P. graminis whereas no DNA fragment was obtained from other non-target wheat fungal pathogens. The detection limit of the primer was 1 ng DNA in a 25-mL PCR reaction. The SSR markers of P. graminis can also be used to detect the presence of latent hyphae in Pgt-infected wheat leaves as early as 30 h post-inoculation. A rapid approach to distinguish P. graminis from similar pathogenic fungi would be anticipated in further study.

The plasmid pGPDGFP under the control of pgpdA promotor was used together with vector pAN7-1 containing the hygromycin resistance cassette to co-transform protoplasts of HG1, Fusarium graminearum from Hubei Province, China. Twelve out of 14 hygromycin-resistant transformants showed green signal under the UV light and contained one or several copies of gfp, as indicated by Southern analysis of genomic DNA digested with different restriction enzymes and hybridized to the gfp probe. A single gfp copy transformant (HG1C5) was selected for further evaluation of 80 Chinese wheat cultivars or advanced lines. The results showed different resistance type to F. graminearum were observed. GFP signals observed in the rachis and adjacent spikes of 70 Chinese wheat lines such as Chuanchongzu 104 indicated both type I (host resistance to the initial infection by the fungus) and type II (resistance to the spread of FHB symptoms within an infected spike) were not observed. While other 10 lines showed type II resistance to F. graminearum with GFP signals only in inoculated spikelets. Development of the mycelium can be intuitively observed and the resistance of wheat to F. graminearum can be identified at 7 days post inoculation (dpi) in this way. The results showed no differences were evaluated between the transformed HG1C5 and the non-transgene artificial inoculation by SAS paired chi-square test and McNemar’s test (P=0.0625).

Methiopyrisulfuron is a novel sulfonylurea herbicide with good activity for annual broadleaf and gramineal weeds control. Present study was to investigate the effects of organic amendments (including peat (PE), sewage sludge (SS), and humic acid (HA)) on adsorption, desorption and leaching of methiopyrisulfuron in soils. The batch equilibration technique was applied for adsorption-desorption experiments and the leaching was tested through soil column simulated experiments under laboratory conditions. The Freundlich model may well describe adsorption-desorption of methiopyrisulfuron on organic amendments, the natural soil, and amended soils. Organic amendments could not only greatly increase the adsorption capacity of methiopyrisulfuron, but also significantly enhance the hysteresis of desorption of methiopyrisulfuron. The correlations between Kf-ads and organic matter content of amended soils were significant, and the correlations between H and soil organic matter in amended soils with PE, SS, and HA were significant too. The results of soil column experiments indicated that organic amendments greatly decreased leaching of methiopyrisulfuron. This study suggested that PE, SS, and HA could greatly influence environmental behavior of methiopyrisulfuron in soils. Use of organic amendments might be an effective management practice for controlling potential pollution of methiopyrisulfuron to environment.

Root border cells (RBCs) originate from the root tip epidermis and surround the root apices. In this study, we evaluated the developmental characteristics and the roles of RBCs in protection of root apices of cucumber and fi gleaf gourd seedlings from CA toxicity. The formation of RBCs and the emergence of the root tip occurred almost simultaneously in root apices of cucumber and fi gleaf gourd seedlings. CA ranging from 0 to 0.25 mol L-1 inhibited root elongation and decreased root cell viability in the root tip, moreover the inhibitory effects of CA were more signifi cant in the CA-sensitive cucumber than in the CA-tolerant fi gleaf gourd. Removal of RBCs from root tips led to more severe CA induced inhibition of root elongation and decline in root cell viability. Increasing CA levels and treatment time decreased the relative viability of attached and detached RBCs. CA also induced a thicker mucilage layer surrounding attached RBCs of both species. Additionally, a signifi cantly higher relative cell viability of attached RBCs and thicker mucilage layers were observed in fi gleaf gourd. These results suggest that RBCs play an important role in protecting root tips from CA toxicity.