A large number of callus from mature seeds of indica rice minghui 63 were obtained through pre-induction on medium with 2 mg/L 2,4-D but without inorganic and organic components for 9 days. Trichosanthin gene was transferred into indica rice minghui 63 by using agrobacterium with the help of bombardment and the transgenic plants were obtained by inducing regeneration. Southern and Western blot analysis showed that the trichosanthin gene had been transferred into genome of minghui 63 and expressed in rice plants. The anti-fungal assay suggested that transgenic rice plants enhanced resistance to infection of Pyricularia oryzae.
Mitosporic Fungi ; drug effects ; Oryza ; genetics ; microbiology ; Plant Diseases ; microbiology ; Plants, Genetically Modified ; Regeneration ; Seeds ; physiology ; Trichosanthin ; genetics

We constructed an expression cassette of the organophosphorus pesticide degrading (opd) gene under the control of the E8 promoter. Then opd was transformed into tomato fruit using an agroinfiltration transient expression system. beta-Glucuronidase (GUS) staining, reverse transcription-polymerase chain reaction (RT-PCR), wavelength scanning, and fluorescent reaction were performed to examine the expression of the opd gene and the hydrolysis activity on coumaphos of organophosphorus hydrolase (OPH) in tomato fruit. The results show that the agroinfiltrated tomato fruit-expressed OPH had the maximum hydrolysis activity of about 11.59 U/mg total soluble protein. These results will allow us to focus on breeding transgenic plants that could not only enhance the degrading capability of fruit and but also hold no negative effects on pest control when spraying organophosphorus pesticides onto the seedlings in fields.
Aryldialkylphosphatase ; genetics ; physiology ; Coumaphos ; pharmacology ; Insecticides ; pharmacology ; Lycopersicon esculentum ; genetics ; metabolism ; Plants, Genetically Modified ; Promoter Regions, Genetic

Two stable transformed lines containing antisense LeETR1 [corrected] or LeETR2 [corrected] sequences and their hybridized line were investigated to determine the effect of LeETR1 [corrected] and LeETR2 [corrected] specificity in the ethylene receptor family in tomato (Lycopersicon esculentum Mill.) on ethylene signaling. The transgenic line ale1 containing antisense LeETR1 [corrected] displayed shorter length of seedling grown in the dark and adult plant in the light, severe epinastic petiole, and accelerated abscission of petiole explant and senescence of flower explant, compared with its wild type B1. The transgenic line ale2 containing antisense LeETR2 [corrected] also exhibited shorter hypocotyls and slightly accelerated abscission. The phenotypes of cross line dale of LeETR1 [corrected] and LeETR2 [corrected] were close to ale1 in many aspects. These results suggested that LeETR1 [corrected] probably plays a relatively important role in ethylene signaling of tomato growth and development.
Ethylenes ; metabolism ; Gene Silencing ; physiology ; Lycopersicon esculentum ; physiology ; Plant Proteins ; genetics ; metabolism ; Plants, Genetically Modified ; physiology ; RNA, Antisense ; physiology ; Receptors, Cell Surface ; genetics ; metabolism

The most nutrients required in the human diet come from plants. The nutritional quality of plant products affects the human healthy. The advance of molecular cloning and transgenic technology has provided a new way to enhance the nutritional value of plant material. Transgenic modification of plant nutritional value has progressed greatly in the following aspects: improving the quality, composition and levels of protein, starch and fatty acid in different crops; increasing the levels of antioxidants (e.g. carotenoids and flavonoids); breeding the new type of plants with medical value for human. To date, many transgenic plants with nutritional enhancement have been developed. These transgenic plant products could be directly used as human diet or as valued materials in developing the "functional food" with especial nutritional quality and healthy effects after they are approved by a series of evaluations on their safety and nutritional efficiency for human being. We designed new zinc finger transcription factors (ZFP-TFs) that can specifically down-regulate the expression of the endogenous soybean FAD2-1 gene which catalyzes oleic acid to linoleic acid. Seed-specific expression of these ZFP-TFs in transgenic soybean somatic embryos repressed FAD2-1 transcription and increased significantly the levels of oleic acid, indicating that the engineered ZFP-TFs are capable of regulating fatty acid metabolism and modulating the expression of endogenous genes in plants.
Antioxidants ; metabolism ; Fatty Acids ; analysis ; biosynthesis ; Humans ; Isoflavones ; Nutritive Value ; Plants ; metabolism ; Plants, Genetically Modified ; genetics ; Transcription Factors ; physiology ; Zinc Fingers

Polyamine is an important physiological regulation substance functioning in a wide variety of biological processes, such as plant growth, development, senescence and adversity stress tolerance, which widely exist in all living organisms. Their biosynthetic pathways have already been revealed, and their physiological roles are being elucidated gradually. Previous work on polyamines biosynthetic deficiency mutants and various transgenic plants facilitates improved understanding of the important roles of polyamines and biosynthetic enzymes in plant growth and development. This paper summarizes researches in the biosynthetic pathways of polyamines in plants, focusing on research advances on functions of genes involved in polyamine metabolism. In addition, the potential research directions, especially the application of the genes in the genetic engineering of plant stress tolerance were also discussed.
Biosynthetic Pathways ; physiology ; Gene Expression Regulation, Plant ; Genes, Plant ; genetics ; Plant Physiological Phenomena ; Plants ; metabolism ; Plants, Genetically Modified ; Polyamines ; metabolism ; Stress, Physiological

An efficient system of genetic transformation and plant regeneration via somatic embryogenesis was established in crownvetch (Coronilla varia L.) by infecting the segments of cotyledons and hypocotyls of 15d-old seedlings with Agrobacterium rhizogenes strain 15834. Hairy roots were produced directly from the wounded surface of the explants or via calluses on hormone-free Murashige and Skoog (MS) medium after infection by A. rhizogenes. Transformed roots grew rapidly either on solid or liquid MS medium, and exhibited typical hairy root phenotypes. The highest transformation frequency (87.4%) was achieved by preculturing cotyledons for 2d and pre-treating the A. rhizogenes with suitable concentration of acetosyringone at logarithmic phase (OD600 = 0.8). The embryogenic calluses with 100% induction frequency were induced from hairy roots on MS medium containing 0.2mg/L 2,4-D, 0.5mg/L NAA and 0.5mg/L KT. Globular-, heart-, torpedo-, and cotyledon shaped somatic embryos were produced orderly and developed into plantlets when transferred the embryogenic calluses on MS medium supplemented with 0.5mg/L KT, 0.2mg/L IBA and 300mg/L proline. The transformed plants did not show differences in morphology except abundant lateral root branches compared to the non-transformed plants. However, the contents of 3-nitropropanic acid in hairy roots and leaves of one of 5 transformed clones were 57.68% and 58.17% in roots and leaves of untransformed plants, respectively. Opine paper electrophoresis revealed the integration and expression of TR-DNA. PCR analysis confirmed that the TL-DNA including 654 bp rol B sequence was inserted into the genome of transformed hairy roots and their regenerated plants.
Fabaceae ; genetics ; growth & development ; physiology ; Plant Roots ; genetics ; growth & development ; physiology ; Plants, Genetically Modified ; genetics ; growth & development ; Regeneration ; Rhizobium ; genetics ; Tissue Culture Techniques ; Transformation, Genetic

OBJECTIVETo evaluate the health effects of parental dietary exposure to GM rice TT51 on the male reproductive system of rat off spring.

METHODSRice-based diets, containing 60% ordinary grocery rice, MingHui63, or TT51 by weight, were given to parental rats (15 males/30 females each group) for 70 days prior mating and throughout pregnancy and lactation. After weaning, eight male offspring rats were randomly selected at each group and fed with diets correspondent to their parents' for 70 days. The effects of exposure to TT51 on male reproductive system of offspring rats were assessed through sperm parameters, testicular function enzyme activities, serum hormones (FSH, LH, and testosterone levels), testis histopathological examination, and the relative expression levels of selected genes along the hypothalamic-pituitary- testicular (HPT) axis.

RESULTSNo significant differences were observed in body weight, food intake, organ/body weights, serum hormone, sperm parameters, testis function enzyme ACP, LDH, and SDH activities, testis histopathological changes, and relative mRNA expression levels of GnRH-R, FSH-R, LH-R, and AR along the HPT axis.

CONCLUSIONThe results of this study demonstrate that parental dietary exposure to TT51 reveals no significant differences on the reproductive system of male offspring rats compared with MingHui63 and control.

Animals ; Diet ; adverse effects ; Female ; Genitalia, Male ; physiology ; Male ; Oryza ; chemistry ; Plants, Genetically Modified ; adverse effects ; chemistry ; Random Allocation ; Rats ; Rats, Wistar

'Shuhui527' is a promising restorer line bred by Rice Research Institute of Sichuan Agricultural University in recent years. However, this line is susceptible to Bacterial Blight (BB), which limits its use. The IRBB60, from the International Rice Research Institute (IRRI), contains dominant genes Xa21 and Xa4 conferring resistance to BB. The objective of this study is to improve the BB resistance of 'Shuhui527' by introgressing Xa21 and Xa4, the two broad-spectrum BB resistance genes, into 'Shuhui527' with IRBB60 as the donor, pTA248 and MP12, linking tightly with Xa21 and Xa4 respectively as DNA markers. BC1 F1 progenies of (Shuhui527 x IRBB60), containing Xa21 and Xa4 identified using PCR screening and with agronomic traits including plant type, grain type and days to heading etc similar to those of 'Shuhui527', were subsequently backcrossed to 'Shuhui527' and self-pollinated to generate BC2 F1 and BC1 F2 . The BC3 F1 and BC3 F2 were subsequently developed using the same approach. Among the 20 BC3 F2 plants, homozygous Xa21 and Xa4,10 plants were the most similar to 'Shuhui527' in the agronomic traits, and were screened using 120 pairs SSR and 100 pairs RAPD markers. Based on the results of the background screening and the performance of the agronomic traits, 5 plants were identified as improved-'Shuhui527' and designated as 527R-5, 527R-6, 527R-8, 527R-9 and 527R-10. The improved-' Shuhui527' lines expressed high resistance to Xanthomonas oryzae pv . oryzae (Xoo) stains C I - C VII, P1 and P6. The evaluation of the polymorphisms and selection accuracies of pTA248 and MP12 demonstrated that the polymorphisms of the two markers were obvious and co-dominant and the accuracies were more than 97% and 83% respectively, indicating the two markers are good for Xa21 and Xa4 in Molecular Marker-assisted Selection.
Genes, Plant ; genetics ; physiology ; Oryza ; genetics ; microbiology ; Plant Diseases ; genetics ; microbiology ; Plants, Genetically Modified ; genetics ; microbiology ; Polymerase Chain Reaction ; Xanthomonas ; pathogenicity

To study the possibilities for improvement of the ornamental character and production of secondary metabolites by using Wedelia trilobata hairy roots, we investigated the induction of W. trilobata L. hairy roots and its consumption changes of carbon resource, nitrogen resource, phosphate and calcium in the medium during liquid culture. The results showed that hairy roots could be incited from the cut edges of leaf explants 7 days after inoculation with Agrobacterium rhizogenes ATCC15834 and could have an autonomous growth on the medium without phytohormones. The PCR amplification showed that rol genes of Ri plasmid of A. rhizogenes was integrated and expressed into the genome of transformed hairy roots. The hairy root line grew very slowly in 0-7 days, very fast from 7 to 21 days. During the liquid culture of hairy roots, sucrose, NO3(-)-N, PO4(3-) and Ca2+ in the medium could be gradually absorbed and utilized with time. The content of NO3(-)-N in the medium was 5.8% of the initial amount at day 7, while sucrose content was about 50% of the initial amount. At day 35, the NO3(-)-N and sucrose content in the medium was 1.82% and 3.39% of the initial amount, respectively. In combination with Ca2+ consumption, PO4(3-) of the medium was rapidly absorbed and utilized. At day 7, the content of PO4(3-) in the spent medium was only 1.76% of the initial amount; but even at day 35, the content of Ca2+ in the spent medium was still 61.3% of the initial amount. The results presented here had provided the possibilities on improvement the ornamental character and how to prepare optimum medium for large scale cultivation and production of secondary metabolites from W. trilobata L. hairy roots.
Culture Techniques ; methods ; Plant Roots ; cytology ; growth & development ; Plants, Genetically Modified ; genetics ; growth & development ; metabolism ; Rhizobium ; genetics ; physiology ; Transformation, Genetic ; Wedelia ; genetics ; growth & development ; microbiology

We describe a genetic transformation method of secondary somatic embryogenesis in alfalfa through cotyledon-stage somatic embryos of alfalfa infected by Agrobacterium strain GV3101. The Agrobacterium strain GV3101 contained binary vector pCAMBIA2301 that had gus gene as reporter and npt II gene as selectable marker. The infected primary embryos were induced through series of medium under 75 mg/L kanamycin selection. We obtained the transgenic alfalfa plants. Then, GUS expression in different tissue of transgenic alfalfa was tested by GUS histochemical analysis. Further, the stable integration and transformation efficiency were tested by polymerase chain reaction and Southern blotting hybridization. The result showed that GUS expression was different in different organs of transgenic alfalfa; the copy number of integrated npt II gene was from 1 to 4; the transformation efficiency via primary somatic embryogenesis was 65.82%.
Agrobacterium ; genetics ; Medicago sativa ; embryology ; genetics ; physiology ; Plant Somatic Embryogenesis Techniques ; Plants, Genetically Modified ; embryology ; genetics ; Tissue Culture Techniques ; Transformation, Genetic