Cytosolic disproportionating enzyme2 is essential for pollen germination and pollen tube elongation in rice.

Degradation of starch accumulated in pollen provides energy and cellular materials for pollen germination and pollen tube elongation. Little is known about the function of cytosolic disproportionating enzyme2 (DPE2) in rice (Oryza sativa). Here, we obtained several DPE2 knockout mutant (dpe2) lines via genomic editing and found that the mutants grew and developed normally but with greatly reduced seed-setting rates. Reciprocal crosses between dpe2 and wild-type plants demonstrated that the mutant was male sterile. In vitro and in vivo examinations revealed that the pollen of the dpe2 mutant developed and matured normally but was defective in germination and elongation. DPE2 deficiency increased maltose content in pollen, whereas it reduced the levels of starch, glucose, fructose, and adenosine triphosphate (ATP). Exogenous supply of glucose or ATP to the germination medium partially rescued the pollen germination defects of dpe2. The expression of cytosolic phosphorylase2 (Pho2) increased significantly in dpe2 pollen. Knockout of Pho2 resulted in a semi-sterile phenotype. We failed to obtain homozygous dpe2 pho2 double mutant lines. Our results demonstrate that maltose catalyzed by DPE2 to glucose is the main energy source for pollen germination and pollen tube elongation, while Pho2 might partially compensate for deficiency of DPE2.

Hypotensive Activity of Transgenic Rice Seed Accumulating Multiple Antihypertensive Peptides.

Peptides derived from food protein have the potential to become antihypertensive agents with relatively few negative side effects. Herein, multiple antihypertensive peptides, extracted from the transgenic rice seed, were administered intragastrically into spontaneously hypertensive rats (SHRs) with different dosages, resulting in a significant decrease in the systolic blood pressure (SBP). Furthermore, for a period of 5 weeks, daily intragastric administration of the transgenic rice flour also significantly reduced the SBP of SHRs but not the Wistar Kyoto normotensive rats (WNRs), most importantly, which did not affect the growth, development, or serum chemistry of SHRs or WNRs and did not cause any pathological changes. Our work provides an alternative source of natural antihypertensive agents.

[Treatment of pediatric malnutrition by acupuncture of Sifeng (EX-UE10) combined with oral administration of Wang's Baochi Pill].

OBJECTIVE: To investigate the clinical effect of acupuncture at Sifeng (EX-UE10) combined with Wang's Baochi Pills in the treatment of pediatric malnutrition, so as to provide a more effective method for pediatric malnutrition. METHODS: A total of 201 children with malnutrition were randomly divided into combined treatment group (n＝102) and control (Baochi Pill) group (n＝99). The children in the combined treatment group were treated by acupuncture stimulation of Sifeng (EX-UE10, till no more yellowish-white effusion out) and oral administration of Wang's Baochi Pills, and those in the control group treated by oral administration of Wang's Baochi Pills only. The course of treatment was one month for both groups. The integral score of symptom was assessed according to the main symptoms as body weight and height and food-intake, and to the secondary symptoms including mentality, agitation, sleep, hair gloss, susceptibility to cold, hydrosis, abdominal distension, and susceptibility to diarrhea or constipation. The therapeutic effect was assessed by consulting the "Criteria for Diagnosis and Therapeutic Effect Evaluation of Syndromes/Illnesses of Traditional Chinese Medicine (TCM)" and "TCM Professional Criteria of the People's Republic of China for Diagnosis and Therapeutic Effect Evaluation of Syndromes/Diseases of TCM Pediatric". RESULTS: After the treatment, the total symptom scores of both groups were significantly decreased in comparison with their own pre-treatment (P<0.05), and the scores of total symptom after the treatment and the 1st, 2nd and 3rd follow-up surveys were obviously lower in the combined treatment group than in the control group (P<0.05). Of the 99 and 102 cases in the control and combined treatment groups, 47 and 59 were cured, 39 and 37 experienced improvement in their symptoms, and 13 and 6 failed, with the effective rate being 86.87% (86/99) and 94.11%(96/102), respectively. The effective rate of the combined treatment was evidently higher than that of the simple medication (P<0.05). CONCLUSION: Acupuncture of Sifeng (EX-UE10) combined with Wang's Baochi Pills is better than administration of Wang's Baochi Pills alone in the therapeutic effect for pediatric malnutrition.

[Syndrome Features of Chinese Medicine in Hyperlipidemia Patients of Han, Uyghur, Kazakh Na- tionalities in Xinjiang Uyghur Autonomous Region].

Objective To observe distribution laws and features of syndrome types of Chinese medicine (CM) in hyperlipidemia patients of Han, Uyghur, Kazakh nationalities in Xinjiang Uyghur Auton- omous Region. Methods Using cluster random sampling, 1 410 hyperlipidemia patients (18 -70 years old ) were recruited from Urumqi, Turpan, Altay, Ili, Aksu, Hetian in Xinjiang Uyghur Autonomous Re- gion. The general condition, susceptible factors, classification of blood lipids, complications, syndromes of CM, tongue figure, etc. clinical data were investigated using self-formulated Epidemiological Investiga- tion Questionnaire on Susceptible Factors in Different Nationalities of Hyperlipemia Patients in Xinjiang (abbreviated as Questionnaire thereafter). Factor analysis and cluster analysis were performed. Results Cronbach's coefficient for the 54 syndrome items in Questionnaire was 0.891, Kaiser-Meyer-Olkin (KMO) 0. 897, Sig <0.05 in Bartlett's sphericity test. Seventeen common factors were obtained using principal component analysis (PCA). Totally 54 common symptoms of hyperlipidemia were screened, which were then divided into 17 groups with 1 -6 symptoms in each group. F4 (soreness and weakness of waist and knees, sour pain in joints and muscles, body numbness, heavy body sensation, cold limbs), F5 (frequent and clear nocturia, dysuria,-dribble of urine, frequent urination at night), F10 (thirsty, no desire for water, tastelessness, hydroadipsia) , F12 (a white complexion with puffiness, hid- ing fever, hypoactive sexual desire) , and F17 (enuresis) were merged as Shen yang deficiency (SYD) ; F2 (fatigue, drowsiness, depression, spiritlessness, fatigue and disinclination to talk) and F15 (poor ap- petite) were merged as Pi-qi deficiency (PQD) ; F3 (dry mouth and dry pharynx, thirsty, propensity for water, bitter mouth, greasy mouth, stingy mouth, irritability and upset) and F16 (dark red tongue proper, greasy tongue fur) were merged as damp-heat trapped in Pi (DHTP). Results of cluster analysis showed that Pi-Shen deficiency (PSD) was most often seen in hyperlipidemia, and main syndrome types were sequenced from high to low as Pi-Shen deficiency type (46. 2%, 652/1 410) , blockage of cardiac vessels type ( 31. 1% , 438/1 410 ), phlegm and blood stasis internal resistance type ( 13. 3% , 187/1 410), Pi-deficiency induced damp abundance type (8. 3%, 123/1 410), Gan-Shen yin deficiency type (0. 7%, 10/1 410). Conclusions Deficiency syndrome was dominant in hyperlipidemia patients of Xinjiang Uyghur Autonomous Region. Phlegm turbidity, damp heat, and etc. were often complicated. The complex situation was manifested to be involved in multiple organs, qi-blood-fluid mixed disease.

Integrated and comparative proteomics of high-oil and high-protein soybean seeds.

We analysed the global protein expression in seeds of a high-oil soybean cultivar (Jiyu 73, JY73) by proteomics. More than 700 protein spots were detected and 363 protein spots were successfully identified. Comparison of the protein profile of JY73 with that of a high-protein cultivar (Zhonghuang 13, ZH13) revealed 40 differentially expressed proteins, including oil synthesis, redox/stress, hydrolysis and storage-related proteins. All redox/stress proteins were less or not expressed in JY73, whereas the expression of the major storage proteins, nitrogen and carbon metabolism-related proteins was higher in ZH13. Biochemical analysis of JY73 revealed that it was in a low oxidation state, with a high content of polyunsaturated fatty acids and vitamin E. Vitamin E was more active than antioxidant enzymes and protected the soybean seed in a lower oxidation state. The characteristics of high oil and high protein in soybean, we revealed, might provide a reference for soybean nutrition and soybean breeding.

Embryo-specific expression of soybean oleosin altered oil body morphogenesis and increased lipid content in transgenic rice seeds.

Oleosin is the most abundant protein in the oil bodies of plant seeds, playing an important role in regulating oil body formation and lipid accumulation. To investigate whether lipid accumulation in transgenic rice seeds depends on the expression level of oleosin, we introduced two soybean oleosin genes encoding 24 kDa proteins into rice under the control of an embryo-specific rice promoter REG-2. Overexpression of soybean oleosin in transgenic rice leads to an increase of seed lipid content up to 36.93 and 46.06 % higher than that of the non-transgenic control, respectively, while the overall fatty acid profiles of triacylglycerols remained unchanged. The overexpression of soybean oleosin in transgenic rice seeds resulted in more numerous and smaller oil bodies compared with wild type, suggesting that an inverse relationship exists between oil body size and the total oleosin level. The increase in lipid content is accompanied by a reduction in the accumulation of total seed protein. Our results suggest that it is possible to increase rice seed oil content for food use and for use as a low-cost feedstock for biodiesel by overexpressing oleosin in rice seeds.

Identification and evaluation of ω-3 fatty acid desaturase genes for hyperfortifying α-linolenic acid in transgenic rice seed.

α-Linolenic acid (ALA) deficiency and a skewed of ω6:ω3 fatty acid ratio in the diet are a major explanation for the prevalence of cardiovascular diseases and inflammatory/autoimmune diseases. There is a need to enhance the ALA content and to reduce the ratio of linoleic acid (LA) to ALA. Six ω-3 (Δ-15) fatty acid desaturase (FAD) genes were cloned from rice and soybean. The subcellular localizations of the proteins were identified. The FAD genes were introduced into rice under the control of an endosperm-specific promoter, GluC, or a Ubi-1 promoter to evaluate their potential in increasing the ALA content in seeds. The ALA contents in the seeds of endoplasmic reticulum (ER)-localized GmFAD3-1 and OsFAD3 overexpression lines increased from 0.36 mg g⁻¹ to 8.57 mg g⁻¹ and 10.06 mg g⁻¹, respectively, which was 23.8- and 27.9-fold higher than that of non-transformants. The trait of high ALA content was stably inheritable over three generations. Homologous OsFAD3 is more active than GmFAD3-1 in catalysing LA conversion to ALA in rice seeds. Overexpression of ER-localized GmFAD3-2/3 and chloroplast-localized OsFAD7/8 had less effect on increasing the ALA content in rice seeds. The GluC promoter is advantageous compared with Ubi-1 in this experimental system. The enhanced ALA was preferentially located at the sn-2 position in triacylglycerols. A meal-size portion of high ALA rice would meet >80% of the daily adult ALA requirement. The ALA-rich rice could be expected to ameliorate much of the global dietary ALA deficiency.