基于broccoli的RNA荧光适体设计与筛选

为了在活细胞RNA成像中提供更加精准的工具,本文旨在基于broccoli的RNA荧光标记创建荧光强度高、稳定性好的绿色荧光适体.采用理性设计对broccoli进行优化,并通过测定光谱特征和理化性质对broccoli变体进行多轮筛选.结果发现基于对RNA荧光适配体broccoli的三轮突变筛选,获得了荧光强度提升3.3倍的broccoli四位点突变体(命名为Bro);再利用设计的3WJ支架,组建了3WJ-n×Bro RNA荧光适配体家族;采用嵌入串联重复Bro的方式可以显著提升适体的荧光强度,其中3WJ-4×Bro的荧光强度超过3WJ-Bro的2.7倍,且表现出更低的盐离子依赖性和更高的热稳定性、光稳定性和监测灵敏度.结果表明本文设计出的3WJ-n×Bro家族并从中筛选出优异的核酸适配体3WJ-4×Bro,可为在RNA水平上实现活细胞RNA成像提供可视化工具.

Selenocysteine methyltransferase SMT catalyzed the synthesis of Se-methylselenocysteine to regulate the accumulation of glucosinolates and sulforaphane in broccoli

Selenocysteine methyltransferase(SMT)is a key enzyme involved in the Se metabolism pathway,and it is responsible for the catalysis of Se-methylselenocysteine(SeMSC)compound formation.Previous studies showed that selenium treatment activated SMT expression and promoted the accumulation of glucosinolates(GSLs)and sulforaphane,but the roles and functional mechanisms of SMT in mediating GSLs and sulforaphane synthesis remain unclear.In this study,we identified the BoSMT gene in broccoli and uncovered its roles in mediating GSLs biosynthesis.Transgenic assays revealed that BoSMT is involved in SeMSC biosynthesis in broccoli.More importantly,the contents of GSLs and sulforaphane were significantly increased in the BoSMT-overexpressing broccoli lines but decreased in the knockdown lines,suggesting that BoSMT played a positive role in regulating GSLs and sulforaphane synthesis.Further evidence indicated that BoSMT-mediated overaccumulation of GSLs and sulforaphane might be due to the increase in the endogenous SeMSC content.Compared with the mock(water)treatment,selenite-induced significantly increases of the SeMSC content in the BoSMT-knockdown plants partially compensated the phenotype of GSLs and sulforaphane loss.Compared with the mock treatment,exogenous SeMSC treatment significantly increased the contents of GSL and sulforaphane and activated GSL synthesis-related gene expression,suggesting that SeMSC acted as a positive regulator for GSL and sulforaphane production.Our findings provided novel insights into selenium-mediated GSLs and sulforaphane accumulation.The genetic manipulation of BoSMT might be a useful strategy for improving the dietary nutritional values of broccoli.

Transcriptomics integrated with metabolomics reveals the mechanism of CaCl_(2)-HCl electrolyzed water-induced glucosinolate biosynthesis in broccoli sprouts

Glucosinolates are important phytochemicals in Brassicaceae.We investigated the effect of CaCl_(2)-HCl electrolyzed water(CHEW)on glucosinolates biosynthesis in broccoli sprouts.The results showed that CHEW treatment significantly decreased reactive oxygen species(ROS)and malondialdeh yde(MDA)contents in broccoli sprouts.On the the 8^(th)day,compared to tap water treatment,the the total glucosinolate content of broccoli sprouts with CHEW treatment increased by 10.6%and calcium content was dramatically enhanced from 14.4 mg/g DW to 22.7 mg/g DW.Comparative transcriptome and metabolome analyses revealed that CHEW treatment activated ROS and calcium signaling transduction pathways in broccoli sprouts and they interacted through MAPK cascades.Besides,CHEW treatment not only promoted the biosynthesis of amino acids,but also enhanced the expression of structural genes in glucosinolate synthesis through transcription factors(MYBs,bHLHs,WRKYs,etc.).The results of this study provided new insights into the regulatory network of glucosinolates biosynthesis in broccoli sprouts under CHEW treatment.

Effects of fresh-cut and storage on glucosinolates profile using broccoli as a case study

Glucosinolates(GLS) contribute to the unique flavour, nutrition, and plant defence of the Cruciferous vegetables. Understanding the GLS changes through postharvest processing is essential for defined preservation. In this study, four different fresh-cut types, whole flower(W),floret(F), quarterly cut floret(QF) and shredded floret(FS) of broccoli, were stored for 0, 1, 2 and 3 day(s) to explore GLS responses to postharvest treatments. As a result, seven GLS were identified, mainly including glucoraphanin(RAA), neoglucobrassicin(NEO), and glucobrassicin(GBC)and accounting for 52.69%, 20.12% and 14.99% of the total GLS(21.92 ± 0.48) μmol · g ^(-1 )DW, respectively. FS had the sharpest decrease in GLS after three days of storage(6.55 ± 0.37) μmol · g-1DW, while QF had the least(10.16 ± 0.33) μmol · g ^(-1 )DW. All GLS components decreased over storage, except for 4-methoxyglucobrassicin(4 ME) in FS and QF, suggesting its key role in serious wound defence. The results suggested certain postharvest approaches influenced the flavour and nutrition of broccoli.

Comparative Study on Biological and Commercial Characteristics of Multiple Varieties of Broccoli(Brassica oleracea L.var.italica Plenck)and Vegetable Soybean(Glycine max(L.)Merr.)

[Objectives]This study was conducted to select vegetable soybean varieties(Glycine max(L.)Merr.)that are suitable for local cultivation and meet export requirements.[Methods]Through continuous years of comparative experiments on broccoli and vegetable soybean varieties,detailed biological characteristic and economic quality data of multiple varieties were obtained.[Results]Vegetable soybean variety Taiwan 75-3 had very prominent early-maturing trait,the highest quality(qualification rate),and higher yield than the control check(CK);and the early-maturing trait of vegetable soybean variety Kaohsiung 9 was also prominent,and its yield was higher than that of the CK.They could be promoted as the main vegetable soybean varieties for spring open field cultivation in this region.Among the tested broccoli varieties,Lake had a higher yield,and was relatively tolerant to cold.It had an early harvest period,and was planted as an early autumn variety in this region.Naihan Youxiu showed the highest yield,good quality,cold resistance,and strong adaptability,making it suitable for planting as a late autumn variety in this region.[Conclusions]This study provides technical guidance for the cultivation of local broccoli and vegetable soybean.

Overexpression of a Broccoli Defensin Gene BoDFN Enhances Downy Mildew Resistance

Plant defensins are small, basic cysteine-rich peptides that play important roles in disease resistance. A gene, designated BoDFN, was isolated from Brassica oleracea var. italica. Gene sequence has been submitted to NCBI with an accession no. of HQ436486. Complete coding sequence of BoDFN is 243 bp in length encoding 80 amino acids. Sequence comparison results showed that BoDFN shared high homology with those of crucifer plants and there were only few DNA base differences. RT-PCR results indicated an increase of gene expression in Hyaloperonospora parasitica infected leaves and revealed a significant increase at 24 and 36 h of treatment. A recombinant plasmid, named pBI121-BoDFN, was constructed and introduced into Agrobacterium tumefacien LBA4404. PCR screening for 65 regenerated plantlets, 17 positive plantlets were obtained, PCR screening results revealed that 17 out of 65 regenerated plantlets were positive. Disease resistant identification results indicated that all positive plants showed an increase in resistance to H. parasitica.

Sulforaphane-rich broccoli sprout extract improves hepatic abnormalities in male subjects

AIM: To evaluate effects of dietary supplementation of sulforaphane(SF)-rich broccoli sprout(BS) extract on hepatic abnormalities in Japanese male participants.METHODS: In a randomized,placebo-controlled,double blind trial,male participants with fatty liver received either BS capsules containing glucoraphanin [GR; a precursor of SF(n = 24)] or placebo(n = 28) for 2 mo. Liver function markers,serum levels of aspartate and alanine aminotransferases(AST and ALT,respectively) and γ-glutamyl transpeptidase(γ-GTP) and an oxidative stress marker,urinary levels of 8-hydroxydeoxyguanosine(8-OHd G),were measured and compared in participants before and after the trial period. In an animal model,chronic liver failure was induced in Sprague-Dawley rats by successive intraperitoneal injection with N-nitrosodimethylamine(NDMA) for 4 wk. Concomitantly,rats received AIN-76 diets supplemented with or without BS extract. Thereafter,rats were sacrificed,and their sera and livers were collected to measure serum liver function markers and hepatic levels of thiobarbituric acid reactive substances(TBARS) levels and hepatic glutathione S-transferase(GST) activity,a prototypical phase 2 antioxidant enzyme.RESULTS: Dietary supplementation with BS extract containing SF precursor GR for 2 mo significantly decreased serum levels of liver function markers,ALT [median(interquartile range),before: 54.0(34.5-79.0) vs after supplementation: 48.5(33.3-65.3) IU/L,P < 0.05] and γ-GTP [before: 51.5(40.8-91.3) vs after: 50.0(37.8-85.3) IU/L,P < 0.05],as well as the alkali phosphatase activity. Placebo showed no significant effects on the markers. The urinary level of 8-OHd G,an established oxidative stress marker,was significantly reduced in participants who had received BS capsules but not the placebo [before: 6.66(5.51-9.03) vs after: 5.49(4.89-6.66) ng/mg-creatinine,P < 0.05]. The reduction of urinary 8-OHd G was significantly correlated with decreased levels of both ALT and γ-GTP [?8-OHd G and ?ALT: Spearman r(r) 0.514 and P = 0.012,?8-OHd G and ?γ-GTP: r = 0.496 and P = 0.016]. Intake of BS extract prevented NDMA-induced chronic liver failure in rats,which was attributable to the suppression of the increase in TBARS through induction of hepatic phase 2 antioxidant enzymes including hepatic GST(86.6 ± 95.2 vs 107.8 ± 7.7 IU/g,P < 0.01).CONCLUSION: Dietary supplementation with BS extract containing the SF precursor GR is likely to be highly effective in improving liver function through reduction of oxidative stress.

Broccoli sprout extract induces detoxification-related gene expression and attenuates acute liver injury

AIM: To investigate the effects of broccoli sprout extract(BSEx) on liver gene expression and acute liver injury in the rat.METHODS: First, the effects of BSEx on liver gene expression were examined. Male rats were divided into two groups. The Control group was fed the AIN-76 diet, and the BSEx group was fed the AIN-76 diet containing BSEx. After a 10-d feeding period, rats were sacrificed and their livers were used for DNA microarray and realtime reverse transcription-polymerase chain reaction(RT-PCR) analyses. Next, the effects of BSEx on acute liver injury were examined. In experiments using acute liver injury models, 1000 mg/kg acetaminophen(APAP) or 350 mg/kg D-galactosamine(D-Gal N) was used to induce injury. These male rats were divided into four groups: Control, BSEx, Inducer(APAP or D-Gal N), and Inducer+BSEx. The feeding regimens were identical for the two analyses. Twenty-four hours following APAP administration via p.o. or D-Gal N administration via i.p., rats were sacrificed to determine serum aspartate transaminase(AST) and alanine transaminase(ALT) levels, hepatic glutathione(GSH) and thiobarbituric acid-reactive substances accumulation and glutathioneS-transferase(GST) activity. RESULTS: Microarray and real-time RT-PCR analyses revealed that BSEx upregulated the expression of genes related to detoxification and glutathione synthesis in normal rat liver. The levels of AST(70.91 ± 15.74 IU/m L vs 5614.41 ± 1997.83 IU/m L, P < 0.05) and ALT(11.78 ± 2.08 IU/m L vs 1297.71 ± 447.33 IU/m L, P < 0.05) were significantly suppressed in the APAP + BSEx group compared with the APAP group. The level of GSH(2.61 ± 0.75 nmol/g tissue vs 1.66 ± 0.59 nmol/g tissue, P < 0.05) and liver GST activity(93.19 ± 16.55 U/g tissue vs 51.90 ± 16.85 U/g tissue, P < 0.05) were significantly increased in the APAP + BSEx group compared with the APAP group. AST(4820.05 ± 3094.93 IU/m L vs 12465.63 ± 3223.97 IU/m L, P < 0.05) and ALT(1808.95 ± 1014.04 IU/m L vs 3936.46 ± 777.52 IU/m L, P < 0.05) levels were significantly suppressed in the D-Gal N + BSEx group compared with the D-Gal N group, but the levels of AST and ALT in the D-Gal N + BSEx group were higher than those in the APAP + BSEx group. The level of GST activity was significantly increased in the D-Gal N + BSEx group compared with the D-Gal N group(98.04 ± 15.75 U/g tissue vs 53.15 ± 8.14 U/g tissue, P < 0.05).CONCLUSION: We demonstrated that BSEx protected the liver from various types of xenobiotic substances through induction of detoxification enzymes and glutathione synthesis.

Antioxidant Activity of Isothiocyanate Extracts from Broccoli

Isothiocyanates(ITCs) extracts were prepared from fresh broccoli.Their antioxidant properties were evaluated by using the in vitro bioassays,including superoxide anion radical( 2 O·-) ,hydroxyl radical(HO·-) and 2,2-diphenyl-1-picrylhydracyl(DPPH·) radical-scavenging methods,lipid peroxidation assay,and reducing power assay.The ITCs extracts exhibited significant dose-dependent antioxidant activities(P<0.01) .Its antioxidant-stability was affected by temperature and storage condition,although it was more stable than vitamin C(Vc) .GC-MS iden-tified sulforaphane in the ITCs extracts.The antioxidant activity of sulforaphane was approximately one tenth to one fifth of that of Vc.Sulforaphane was experimented to be one of dominating contributors to antioxidant capacity of ITCs extracts.However,there was no direct antioxidant activity found for benzyl isothiocyanate(BITC) in vitro. Therefore,it could be concluded that the general group(-N=C=S) was not the essential part for the antioxidant activity of sulforaphane.It is suggested that the ITCs extracts from broccoli can be developed into a promising food additive for healthy food.

Effect of Different Growth State of Broccoli (Brassica oleracea var. italica) on Low Temperature Induction

In order to study the responding of different growth state on low temperature in Broccoli (Brassica oleracea var. italica), we took prematurity broccoli hybrid as the objects. It was found that growth state was varied under different sowing time , moreover, stem became wider with the increasing of light density at the same leaf age. Seedling age of responding to low temperature vernalization only when they grew five leaves above and with over 3.03 ±0.07 centimeter stalk width in 'Qingfeng Broccoli 103'. The older leaf age was and the stronger plant was, the more sensitive and the shorter demanded duration time on low temperature was, and the shorter time required when the plants entered into critical period of floral bud differentiation.

Genetic diversity and population structure analysis of 161 broccoli cultivars based on SNP markers

To better understand the genetic diversity and population structure of broccoli cultivars planted in China,a total of 161 representative broccoli cultivars in the past 25 years were collected and analysed based on single nucleotide polymorphism(SNP)markers.Ten pairs of primers with good polymorphism and high resolution were screened from 315 pairs of SNP primers by 3 broccoli accessions(inbred lines)with different phenotypes and maturity.The 10 pairs of SNP primers were selected,producing 78 alleles.The diversity analysis indicated that the polymorphism information content(PIC)of SNP primer ranged from 0.64 to 0.90.The observed number of alleles(Na)was 2.00,the effective number of alleles(Ne)was 1.11–2.00,the Nei’s gene diversity(H)was 0.10–0.50,and Shannon information index(I)was 0.20–0.70 using PopGene32 software.The clustering results showed that the 161 broccoli cultivars could be divided into 4 major subgroups(A,B,C and D),foreign cultivars were all assigned to subgroup A,and domestic cultivars were assigned to 3 subgroups of B,C,and D.This study indicated that some domestic cultivars and foreign cultivars were similar in genetic background,but most domestic cultivars were still different from the Japanese cultivars.When K=2,the population structure result presented that 161 broccoli cultivars could be divided into 1 simple group(2 groups)and 1 mixed group.When Q≥0.6,143(88.82%)broccoli cultivars belonged to the simple groups.In simple groups 68(42.24%)broccoli cultivars of group 1 were derived from Japan,the United States,Switzerland,the Netherlands,China-Taiwan,and China-Mainland;75(46.58%)broccoli cultivars belonged to group 2;when Q<0.6,18(11.18%)broccoli cultivars belonged to the mixed groups.This study is helpful to understand the diversity and resolution of broccoli cultivars from worldwide,which is beneficial to plant breeding and materials innovation.And meanwhile,this result is also used for construction of broccoli fingerprint serving for cultivar identification.

Effects of 6-Benzylaminopurine–Calcium Chloride–Salicylic Acid on Yellowing and Reactive Oxygen Metabolism of Broccoli

In this study,we investigated the effects of 6-benzylaminopurine(6-BA)–calcium chloride(CaCl_2)–salicylic acid(SA) treatment on the yellowing and reactive oxygen metabolism of harvested broccoli heads. We dipped fresh broccoli heads in a compound solution(0.6 mmol/L 6-BA + 40 mmol/L CaCl_2 + 3 mmol/L SA) for 5 min and then stored them at 23 °C for 4 days. The results showed that the 6-BA–CaCl_2 –SA postharvest treatment effectively retarded the increase in color values(e.g.,variations from black to white,from green to red,and from blue to yellow) and the decline in chlorophyll content of the broccoli heads. Compared with the control broccoli,the rate of superoxide anion radical(O_2^( ·-)) production and the hydrogen peroxide(H_2O_2) content were lowered by the treatment. We also found significant diff erences in the activities of superoxide dismutase(SOD),catalase(CAT) and peroxidase(POD) in the treated broccoli. Based on these results,we consider 6-BA–CaCl_2 –SA to inhibit the accumulation of reactive oxygen,delay the degradation of chlorophyll,and prolong the shelf life of broccoli heads at 23 °C.

On Spatial Distribution Pattern of Broccoli Clubroot Disease and and the Sampling Technique

[Objective] To further improve the prediction and forecast and continuous control ability of broccoli clubroot disease. [Methods] The spatial distribution pattern of diseased or infected plants was analyzed using the least square method, fre- quency distribution, aggregation index, m＊-m regression analysis and Taylor＇s pow- er law model. [Result] The field distribution of broccoli plants with clubroot disease tended to be aggregated distribution, m＇-m regression analysis showed that the el- ementary composition of the spatial distribution of diseased or infected plants was individual colony, the individuals attracted each other; the disease had obvious dis- ease focus in the field, and the individual colony showed uniform distribution pattern in the field. Taylor＇s power law showed that the spatial pattern of individual dis- eased or infected plant with clubroot disease tended to be uniform distribution with the increase of the density. On the basis of this, Iwao optimal theoretical sampling model and sequential sampling model were established, namely N =273.954 1/m- 59.698 5, To （N）=0.368 4N±1.926 8√N, respectively, it meant that when surveying N plants, if the accumulative incidence rate exceeded upper bound, the field can be set as control object; if the accumulative incidence rate didn＇t reach lower bound, it can be set as uncontrol field; if the accumulative incidence rate was between upper bound and lower bound, it should be surveyed continuously until the maximum sample size （mo=0.368 4） appeared, that was, the disease incidence was 15%, so the sampling number should be 684 plants. [Conclusion] The research results had very important instructive meaning for disease control.

Enhanced resistance to Botrytis cinerea and Rhizoctonia solani in transgenic broccoli with a Trichoderma viride endochitinase gene

A endochitinase gene（Tch） from the fungus Trichoderma viride was introduced into broccoli（Brassica oleracea var. italica） by Agrobacterium-mediated transformation. Sixty-eight putative transformants were obtained and the presence of the Tch gene was confirmed by both PCR and Southern blot analysis. RT-PCR analysis showed an accumulation of the transcript encoding the endochitinase protein in the transgenic plants. Using real-time quantitative PCR, the expression profiling of endochitinase gene was analyzed. Primary transformants and selfed progeny were examined for expression of the endochitinase using a fluorometric assay and for their resistance to the pathogenic fungi Botrytis cinerea and Rhizoctonia solani. The endochitinase activities in T0 in vitro plants, T0 mature plants and T1 mature plants were correlated with leaf lesions, and the transgenic line T618 had high endochitinse activities of 102.68, 114.53 and 120.27 nmol L–1 MU min–1 mg–1 protein in the three kinds of plants, respectively. The endochitinase activity showed a positive correlation with the resistance to the pathogens. Most transgenic T0 broccoli had increased resistance to the pathogens of B. cinerea and R. solani in leaf assays and this resistance was confirmed to be inheritable. These findings suggested that expression of the Tch gene from T. viride could enhance resistance to pathogenic fungi in Brassica species.

Effects of broccoli extract and various essential oils on intestinal and faecal microflora and on xenobiotic enzymes and the antioxidant system of piglets

Objective: Since the ban of antibiotics as growth promoting feed additives in the EU in 2006 research in alternatives has gained importance. Phytogenic feed additives represent a heterogenous class of different plant derived substances that are discussed to improve the health of farm animals by direct and indirect antioxidant effects and by influencing microbial eubiosis in the gastrointestinal tract. Consequently our study aimed to investigate the influence of broccoli extract and the essential oils of tur- meric, oregano, thyme and rosemary, as selected individual additives, on intestinal and faecal microflora, on xenobiotic enzymes, and on the antioxidant system of piglets. Methods: 48 four weeks old male weaned piglets were assigned to 6 groups of 8. The piglets were housed individually in stainless steel pens with slatted floor. The control group (Con) was fed a diet without an additive for 4 weeks. The diet of group BE contained 0.15 g/kg sulforaphane in form of a broccoli extract. 535, 282, 373 and 476 mg/kg of the essential oils of turmeric (Cuo), oregano (Oo), thyme (To) and rosemary (Ro) were added to the diets of the remaining 4 groups to stan-dardise supplementation to 150 mg/kg of the oils’ key terpene compounds ar-turmerone, carvacrol, thymol and 1,8-cineole. The composition of bacterial microflora was examined by cultivating samples of jejeunal and colonic mucosa and of faeces under specific conditions. The mRNA expression of xenobiotic and antioxidant enzymes was determined by reversing transcrip- tase real time detection PCR (RT-PCR). Total antioxidant status was assayed using the Trolox Equivalent Antioxidant Capacity (TEAC), and lipid peroxidation was determined by measuring thiobarbioturic acid reactive substances (TBA- RS). Results: Compared to Con piglets all additives positively influenced weight gain and feed conversion in week 1. Over the whole trial period no significant differences in performance parameters existed between the experimental groups. Compared to group Con performance of Ro piglets was, however, slightly impaired. Com- pared to Con piglets Cuo, Oo and To increased the ratio of Lactobacilli:E. coli attached to the jejunal mucosa, whereas BE and Ro impaired this ratio slightly. In contrast in colonic mucosa Ro improved Lactobacilli:E. coli ratio. In faecal samples an improvement of Lactobacilli:E. coli ratio could be analysed for To and Ro. Ro was the only additive that reduced the incidence rate of piglets tested positive for enterotoxic E. coli (ETEC). All additives significantly increased jejunal TEAC and reduced TBA-RS. In the liver BE, Cuo, Oo and To increased TEAC in tendency and Ro significantly. Liver TBA-RS were slightly reduced by all additives compared to Con piglets. Whereas the influence of BE, To and Ro on jejunal TEAC mainly was derived from the induction of xenobiotic and antioxidant enzymes (indirect antioxidant effects), Cuo and Oo influenced TEAC by direct antioxidant effects. Discussion and Conclusions: Our results have shown: That within the labiatae oils Oo and To have the potential to improve performance slightly. That phytogenic substances have a small but not sig- nificant influence on intestinal microflora. That phytogenic feed additives up-regulate the anti- oxidant system of piglets either by direct or by indirect antioxidant effects and that they may thereby improve health status. That within the labiatae oils Oo has a high direct antioxidant potential whereas Ro potently induces xenobiotic and antioxidant enzymes. That broccoli extract is an attractive new phytogenic additive, improving antioxidant status by indirect antioxidant effects. That defined combinations of selected phytogenic substances may produce additive effects. That health promoting effects of phytogenic additives in the future should be studied systematically under the challenge with pathogenic microorganisms or food derived to-xins.

<i>In vitro</i>Bile Acid Binding of Mustard Greens, Kale, Broccoli, Cabbage and Green Bell Pepper Improves with Sautéing Compared with Raw or Other Methods of Preparation

Bile acid binding potential of foods and food fractions has been related to lowering the risk of heart disease and that of cancer. Steam cooking has been observed to significantly improve bile acid binding of green/leafy vegetables. It was hypothesized that other cooking methods could further improve the bile acid binding of various vegetables. Sautée cooking resulted in in vitro bile acid binding measured on a dry matter basis relative to cholestyramine of 14% for mustard greens and kale, 9% for broccoli, 8% for collard greens, 6% for cabbage, and 5% for green bell pepper. These results point to the significantly different (P ≤ 0.05) health promoting potential of mustard greens = kale > broccoli > collard greens > cabbage > green bell pepper. Sautéing significantly improved in vitro bile acid binding of mustard greens, kale, broccoli, cabbage and green bell pepper compared with steaming, boiling or raw (uncooked). Collard greens exhibited significantly higher bile acid binding by steaming compared with sautéing, boiling or raw. Data suggest that the cooking method with most heath promoting potential for mustard greens, kale, broccoli, cabbage and green bell pepper should be sautéing. Steaming should be used for collard greens as the cooking method. These green/leafy vegetables, when consumed regularly after sautéing, would promote a healthy lifestyle and have the potential to lower the risk of premature degenerative diseases.

Effects of Humic Acid and PASP Compound Fertilizer Application on Growth,Quality and Yield of Broccoli

[Objective] This study was conducted to analyze the effects of humic acid and PASP on the growth of broccoli to provide a basis for fertilizer application.[Method] A field experiment was carried out to investigate broccoli growth, yield and quality by using humic acids and PASP as fertilizer synergists in the mixture with compound fertilizer. [Results] The treatment of 5‰ humic acid ＋ 3‰ PASP ＋ compound fertilizer exhibited the plant height, stem diameter and yield increased by8.3%, 13.3% and 10.3%, respectively, and thus was the best; and the treatment showed a single curd weight of 478 g and a commodity rate of 89.4%, and had lower economic cost than treatment of 10‰ humic acid ＋ 6‰ PASP ＋ compound fertilizer. [Conclusion] In summary, 5‰ humic acid ＋ 3‰ PASP ＋ compound fertilizer is suitable for wide application in broccoli culture.

The protoplasts isolation,culture and shoots regeneration of broccoli(Brassica oleracea var.italica)

Eight F1-hybrid cultivars of broccoli were studied.We obtained cell division,celled colonies and p-calli in 5 cultivars,roots and shoots regeneration in one cultivar.The leavesof propagated plantlets in vitro were cut into 1—2mm pieces,isolated with an enzyme solutioncontaining 2% cellulase and 1%macerase on a rotary shaker（50 rpm,21℃,3h,2500 lux light）,and purified with a 0.5M sucrose solution.The purified protoplasts were placed on a drop of 1%agarose.2—3 ml liquid medium was added around the agarose drops,and all of the cultures wereincubated at 25℃ under light（4000 lux）for 16 hours.3—5 days after isolation the cell divisionwas found.About 7 days after incubation 4 multicellular colonies were formed.After 3—5 wksome p-calli were developed.When the p-calli were 2—3 mm in diameter it was transferred to asolidified medium.Once they were developed to 1 cm in diameter they were transferred on a re-generation medium.About 5 months after incubation some roots and shoots grown from the calliwere

Broccoli sprouts juice prevents lens protein aggregation in streptozotocin-induced diabetic rat

AIM: To investigate the effect of broccoli sprouts juice in preventing lens protein aggregation in diabetic rat model.METHODS: Totally 25 male Wistar rats were divided into negative control group, diabetic group without juice treatment as positive control and diabetic group given broccoli sprouts juice 0.25, 0.5 and 1 g/d for 28 d. Diabetic rat model was obtained by administering a single dose streptozotocin 65 mg/kg intraperitoneal. At the end of the study, all rats were examined for fasting blood sugar level(FBS), visual cataract score using slit lamp biomicroscope and lens αB-crystallin expression using Western blot method. Statistical analysis was performed using one way ANOVA with post hoc test, correlation test, and simple linear regression.RESULTS: Positive control group had the highest cataract score and lens aggregated αB-crystallin expression. Broccoli sprout juice dose of 1 g/d group had the mildest cataract score, as well as the expression of lens aggregated αB-crystallin compared to treatments groups 1 and 2, opposite to lens native αB-crystallin expression. The broccoli sprout juice groups gave a significant decrease in cataract score, and also in lens aggregated αB-crystallin expression in diabetic rat models(P<0.05). CONCLUSION: Broccoli sprout juice has a significant effect in preventing lens protein aggregation in diabetic rat model. The higher dose gives better visual cataract scores, lower lens aggregated αB-crystallin expression and higher lens native αB-crystallin expression.

Effect of Hot Water Treatment on Postharvest Shelf Life and Quality of Broccoli

Broccoli was stored at 0, 10, or 20℃after immersion in hot water (38 - 52℃) for 10 or 30 min. Yellowing of broccoli was significantly slowed and shelf life significantly increased when broccoli was treated with hot water at 42 - 46℃and then stored at 10 or 20℃. Heat injury occurred when treatment was higher than 46℃in some varieties. Broccoli lasted 2-3 days longer when stored at 10℃ and 1-2 days longer when stored at 20℃ after hot water treatment at 46℃. There was no significant effect of treatment on shelf life after long time storage at 0℃. Weight loss was reduced by hot water treatment and the respiration behavior of the broccoli was also changed.