Papaya seed represents a rich source of biologically active isothiocyanate

In the present study, papaya (Carica papaya) seed and edible pulp were carefully separated and then the contents of benzyl isothiocyanate and the corresponding glucosinolate (benzyl glucosinolate, glucotropaeolin) quantified in each part. The papaya seed with myrosinase inactivation contained >1 mmol of benzyl glucosinolate in 100 g of fresh seed. This content is equivalent to that of Karami daikon (the hottest Japanese white radish) or that of cress. The papaya seed extract also showed a very high activity of myrosinase and, without myrosinase inactivation, produced 460 micromol of benzyl isothiocyanate in 100 g of seed. In contrast, papaya pulp contained an undetectable amount of benzyl glucosinolate and showed no significant myrosinase activity. The n-hexane extract of the papaya seed homogenate was highly effective in inhibiting superoxide generation and apoptosis induction in HL-60 cells, the activities of which are comparable to those of authentic benzyl isothiocyanate.     

Bioactive compounds, myrosinase activity, and antioxidant capacity of white cabbages grown in different locations of Spain

The influence of two Spanish growing locations with well-differentiated climatic conditions (northern and eastern areas) on the main bioactive compounds, glucosinolates (GLS), total phenolic compounds (TPC), and vitamin C, as well as myrosinase activity and antioxidant capacity in five white cabbage ( Brassica oleracea L. var. capitata) cultivars was investigated. Cabbages with the highest concentration of total GLS presented the highest vitamin C level (r = 0.75, P ≤ 0.05) and the lowest antioxidant capacity (r = -0.76, P ≤ 0.05). The cultivars with the highest vitamin C content had the lowest myrosinase activity (r = -0.89, P ≤ 0.05) and antioxidant capacity (r = -0.86, P ≤ 0.05), whereas those with the largest TPC amount showed the highest antioxidant capacity (r = 0.71, P ≤ 0.05). Cabbage cultivars grown in the northern area of Spain with low temperatures and radiation led to higher mean values of myrosinase activity (29.25 U/g dm), TPC (10.0 GAE mg/g dm), and antioxidant capacity (81.6 μmol Trolox/g dm), whereas cultivars grown in the eastern area with high temperature and radiation led to larger mean values of GLS (14.3 μmol/g dm) and vitamin C (5.3 mg/g dm). The results of this investigation provide information regarding the most suitable Spanish growing location to produce white cabbage with an optimized content of health-promoting compounds.     

Effect of methyl jasmonate on phenolic compounds and carotenoids of romaine lettuce (Lactuca sativa L.)

The effect of exogenous methyl jasmonate (MeJA) on antioxidative compounds of romaine lettuce ( Lactuca sativa L.) was investigated. Lettuces were treated with various MeJA solutions (0, 0.05, 0.1, 0.25, and 0.5 mM) before harvest. Total phenolic compounds content and antioxidant capacity of romaine lettuce significantly increased after MeJA treatments (0.1, 0.25, and 0.5 mM). The total content of phenolic compounds of the romaine lettuce treated with 0.5 mM MeJA (31.6 microg of gallic acid equivalents/mg of dry weight) was 35% higher than that of the control. The increase in phenolic compound content was attributed to a caffeic acid derivative and an unknown phenolic compound, which also contributed to increased antioxidant capacity. The induction of phenylalanine ammonia-lyase (PAL) activity by the MeJA treatment indicated that phenolic compounds were altered due to the activation of the phenylpropandoid pathway. Total content of carotenoids, including lutein and beta-carotene, of the MeJA-treated lettuce did not change after 8 days of treatment, whereas the content of the control without MeJA decreased after 8 days. This research indicated that preharvest application of MeJA could increase the nutritional value of romaine lettuce under determined conditions discussed in this work.     

Comparison of the glucosinolate-myrosinase systems among daikon (Raphanus sativus, Japanese white radish) varieties

Myrosinase is a cytosolic plant enzyme present in daikon ( Raphanus sativus, Japanese white radish) roots that hydrolyzes 4-methylthio-3-butenyl glucosinolate (MTBGLS) into the natural pungent agent 4-methylthio-3-butenyl isothiocyanate (MTBITC), which possesses antimicrobial, antimutagenic, and anticarcinogenic properties. The concentration of MTBGLS, myrosinase activity, and production of MTBITC in seven daikon varieties (one conventional and six heirlooms) were determined to rank the activity of the glucosinolate-myrosinase system and identify critical factors influencing the production of MTBITC. The six heirloom varieties produced 2.0-11.5 times higher levels of MTBITC as compared to the conventional variety, Aokubi, which is consumed by the present Japanese population. The myrosinase was located exclusively in the outer epidermal layer in Aokubi, and MTBGLS was widely distributed throughout the root tissue. Although the skin is a potentially rich source of myrosinase in Aokubi, the skin is usually peeled off in the current practice of preparing daikon for cooking. New practices are therefore proposed for the preparation of daikon tubers that eliminate the peeling of the skin to avoid removing the enzyme needed to convert MTBGLS to the health-beneficial MTBITC. It is also concluded that the consumption of heirloom daikon varieties in addition to changes in food preparation will optimize the health benefits of daikon.     

Non-pathogenic Fusarium strains protect the seedlings of Lepidium sativum from Pythium ultimum

Two root-colonizing Fusarium strains, Ls-F-in-4-1 and Rs-F-in-11, isolated from roots of Brassicaceae plants, induced the resistance in Lepidium sativum seedlings against Pythium ultimum. These strains caused an increase in the content of benzyl isothiocyanate, and of its precursor glucotropaeolin, in the roots of the host plants. The increased isothiocyanate content is one of the factors contributing to the resistance of L. sativum against P. ultimum. To be transformed into the fungitoxic compound benzyl isothiocyanate, glucotropaeolin has to be hydrolyzed by myrosinase, which can be produced either by plants or microorganisms. The Fusarium strain Ls-F-in-4-1 has a myrosinase activity but the strain Rs-F-in-11 has not. These results suggest that both strains are able to trigger the metabolic pathway leading to benzyl isothiocyanate production in the plant. In the case of the myrosinase-negative strain Rs-F-in-11, hydrolyzation into isothiocyanate is only due to the myrosinase activity of the plant, and in the other case, the myrosinase produced by the strain Ls-F-in-11 also would contribute to the production of isothiocyanate. This paper reports a new mode of action of non-pathogenic Fusarium strains in controlling P. ultimum.     

Aqueous extract from Spanish black radish (Raphanus sativus L. Var. niger) induces detoxification enzymes in the HepG2 human hepatoma cell line

Spanish black radish (Raphanus sativus L. var. niger) is a member of the Cruciferae family that also contains broccoli and Brussels sprouts, well-known to contain health-promoting constituents. Spanish black radishes (SBR) contain high concentrations of a glucosinolate unique to the radish family, glucoraphasatin, which represents >65% of the total glucosinolates present in SBR. The metabolites of glucosinolates, such as isothiocyanates, are implicated in health promotion, although it is unclear whether glucosinolates themselves elicit a similar response. The crude aqueous extract from 0.3 to 3 mg of dry SBR material increased the activity of the phase II detoxification enzyme quinone reductase in the human hepatoma HepG2 cell line with a maximal effect at a concentration of 1 mg/mL. Treatment of HepG2 cells with the crude aqueous extract of 1 mg of SBR per mL also significantly induced the expression of mRNA corresponding to the phase I detoxification enzymes: cytochrome P450 (CYP) 1A1, CYP1A2, and CYP1B1 as well as the phase II detoxification enzymes: quinone reductase, heme oxygenase 1, and thioredoxin reductase 1. Previous studies have shown that the myrosinase metabolites of different glucosinolates vary in their ability to induce detoxification enzymes. Here, we show that while glucoraphasatin addition was ineffective, the isothiocyanate metabolite of glucoraphasatin, 4-methylthio-3-butenyl isothiocyanate (MIBITC), significantly induced phase II detoxification enzymes at a concentration of 10 microM. These data demonstrate that the crude aqueous extract of SBR and the isothiocyanate metabolite of glucoraphasatin, MIBITC, are potent inducers of detoxification enzymes in the HepG2 cell line.     

Kinetics of the stability of broccoli (Brassica oleracea Cv. Italica) myrosinase and isothiocyanates in broccoli juice during pressure/temperature treatments

The Brassicaceae plant family contains high concentrations of glucosinolates, which can be hydrolyzed by myrosinase yielding products having an anticarcinogenic activity. The pressure and temperature stabilities of endogenous broccoli myrosinase, as well as of the synthetic isothiocyanates sulforaphane and phenylethyl isothiocyanate, were studied in broccoli juice on a kinetic basis. At atmospheric pressure, kinetics of thermal (45-60 degrees C) myrosinase inactivation could be described by a consecutive step model. In contrast, only one phase of myrosinase inactivation was observed at elevated pressure (100-600 MPa) combined with temperatures from 10 up to 60 degrees C, indicating inactivation according to first-order kinetics. An antagonistic effect of pressure (up to 200 MPa) on thermal inactivation (50 degrees C and above) of myrosinase was observed indicating that pressure retarded the thermal inactivation. The kinetic parameters of myrosinase inactivation were described as inactivation rate constants (k values), activation energy (Ea values), and activation volume (Va values). On the basis of the kinetic data, a mathematical model describing the pressure and temperature dependence of myrosinase inactivation rate constants was constructed. The stability of isothiocyanates was studied at atmospheric pressure in the temperature range from 60 to 90 degrees C and at elevated pressures in the combined pressure-temperature range from 600 to 800 MPa and from 30 to 60 degrees C. It was found that isothiocyanates were relatively thermolabile and pressure stable. The kinetics of HP/T isothiocyanate degradation could be adequately described by a first-order kinetic model. The obtained kinetic information can be used for process evaluation and optimization to increase the health effect of Brassicaceae.     

Metabolomic analysis of phenolic compounds in buckwheat (Fagopyrum esculentum M.) sprouts treated with methyl jasmonate

The effects of exogenous methyl jasmonate (MeJA) on phytochemical production in buckwheat sprouts cultivated under dark conditions (0, 1, 3, 5, and 7 d) were investigated by metabolomic analysis, using ultra performance liquid chromatography-quadrupole-time-of-flight (UPLC-Q-TOF) mass spectroscopy (MS) and partial least-squares-discriminant analysis (PLS-DA). MeJA-treated and control groups showed no differences in growth but were clearly discriminated from each other on PLS-DA score plots. The metabolites contributing to the discrimination were assigned as chlorogenic acid, catechin, isoorientin, orientin, rutin, vitexin, and quercitrin, which have various health effects. Moreover, isoorientin, orientin, rutin, and vitexin were assigned as the main phytochemicals of sprouts cultivated under dark conditions. The accumulation of these metabolites caused the phenolic compound content and antioxidant activity of the sprouts to increase. Further, this study revealed that their accumulation resulted from the stimulation of the phenylpropanoid pathway by MeJA treatment. Therefore, these metabolites may be useful for better understanding the effects of MeJA on buckwheat sprout phytochemicals and contribute to improving the functional quality of the sprouts.     

Changes in glucosinolate concentrations, myrosinase activity, and production of metabolites of glucosinolates in cabbage (Brassica oleracea Var. capitata) cooked for different durations

In cabbage, glucosinolates such as sinigrin are hydrolyzed by plant myrosinase to allyl isothiocyanate (AITC), allyl cyanide, and, in the presence of an epithiospecifier protein, 1-cyano-2,3-epithiopropane (CEP). Isothiocyanates have been implicated in the cancer-protective effects of Brassica vegetables. The effect of processing on the hydrolysis of glucosinolates was investigated in cabbage. Cabbage was steamed or microwaved for six time durations over 7 min. Glucosinolate concentrations were slightly reduced after microwave cooking (P < 0.001) but were not influenced after steaming (P < 0.05). Myrosinase activity was effectively lost after 2 min of microwave cooking and after 7 min of steaming. Hydrolysis of residual glucosinolates following cooking yielded predominantly CEP at short cooking durations and AITC at longer durations until myrosinase activity was lost. Lightly cooked cabbage produced the highest yield of AITC on hydrolysis in vitro, suggesting that cooking Brassica vegetables for a relatively short duration may be desirable from a health perspective.     

Effect of methyl jasmonate on secondary metabolites of sweet basil (Ocimum basilicum L.)

The effect of methyl jasmonate (MeJA) in terms of its induction of inherent bioactive chemicals in sweet basil (Ocimum basilicum L.) was evaluated after MeJA was sprayed on healthy basil plants. The total phenolic content of the sweet basil significantly increased after 0.1 and 0.5 mM MeJA treatments compared with the control not subjected to MeJA. Two phenolic compounds, rosmarinic acid (RA) and caffeic acid (CA), were identified as strong antioxidant constituents of the sweet basil. Their amounts also significantly increased after the MeJA treatment. In addition, eugenol and linalool increased 56 and 43%, respectively, by the 0.5 mM MeJA treatment. Due to the accumulation of RA, CA, and eugenol, which possess strong 2,2-diphenyl-1-picrylhydrazyl (DPPH*) free radical scavenging activities, the antioxidant activity of the sweet basil extract was 2.3-fold greater than that of the control after the 0.5 mM MeJA treatment. In the DPPH* assay, the EC50 values of RA, CA, and eugenol were determined as 23, 46, and 59 microM, respectively, which indicated they were 6-, 3-, and 2.4-fold more efficient than BHT (140 microM). Besides, an unidentified HPLC peak in the methanolic extract of the sweet basil was 4.3-fold higher than that of the control after the 0.5 mM MeJA treatment.     

6-苄氨基嘌呤处理对鲜切西兰花硫代葡萄糖苷代谢的影响

为探讨6-苄氨基嘌呤（6-Benzylaminopurine,6-BA）处理对鲜切西兰花中硫代葡萄糖苷（简称硫苷）代谢的作用机理,该研究首先鉴定得出西兰花花球中分别以萝卜硫苷和萝卜硫素为主的12种硫苷和6种水解产物,然后以30 mg/L的6-BA溶液对鲜切西兰花进行处理,研究外源6-BA处理对其硫苷含量、黑芥子酶活性、硫苷水解产物及其代谢途径关键基因表达水平等的影响。结果表明,6-BA处理通过延缓西兰花花球叶绿素含量下降从而延迟其黄化进程;同时,显著提高了脂肪族硫苷合成相关基因MYB28、CYP79F1、CYP83A1、ST5b、FMOGS-OX1,和吲哚族硫苷合成关键基因MYB51、CYP79B2及CYP83B1的表达水平（P<0.05）,其中6-BA处理组FMOGS-OX1的表达量在贮藏第6天时为对照组的1.93倍,MYB51的表达量为对照组1.41～1.91倍,CYP79B2的表达量在4 d后高出对照组41.90%～93.75%;由此,6-BA处理保持了其组织较高的萝卜硫苷、3-甲基亚磺酰丙基硫苷、吲哚甲基硫苷和4-甲氧基吲哚甲基硫苷的含量,总硫苷含量为对照组的2.46～4.79倍;此外,6-BA处理的西兰花花球组织黑芥子酶活性和其基因MYR的表达水平在4～8 d期间分别高出对照组15.32%～90.22%和25.86%～33.73%,并显著提高了其组织萝卜硫素等异硫氰酸酯水解产物的含量（P<0.05）,其中萝卜硫素含量高于对照组17.58%～25.39%,但对硫苷水解关键基因ESP的表达量并无显著影响（P>0.05）。因此,6-BA处理可通过提高硫苷合成相关基因和水解关键基因来保持鲜切西兰花花球中硫苷的含量和异硫氰酸酯的水平,为西兰花功能活性成分的保持提供理论和技术支持。     

Comparative study of the effects of solid-state fermentation with three filamentous fungi on the total phenolics content (TPC), flavonoids, and antioxidant activities of subfractions from oats (Avena sativa L.)

The aim of present work was to investigate the effect of solid-state fermentation with filamentous fungi (Aspergillus oryzae var. effuses, Aspergillus oryzae, and Aspergillus niger) on total phenolics content (TPC), flavonoids, and antioxidant activities of four subfractions of oat, namely, n-hexane, ethyl acetate (EA), n-butanol, and water, and compare them to their corresponding subfractions of unfermented oat. The TPC and total flavonoids increased dramatically, especially in EA subfractions (p < 0.05). The levels of antioxidant activity of subfractions were also significantly enhanced (p < 0.05). The highest antioxidant activities were also found in the EA subfractions. The polyphenols in EA were analyzed by high-performance liquid chromatography at 280 nm. Most polyphenols were increased remarkably, especially ferulic and caffeic acids. There was a clear correlation between the TPC and antioxidant activity. In conclusion, fungi fermentation is a potential bioprocess for increasing the TPC, flavonoids, and antioxidant activities of oat-based food.     

Effect of roasting on phenolic content and antioxidant activities of whole cashew nuts, kernels, and testa

The effect of roasting on the content of phenolic compounds and antioxidant properties of cashew nuts and testa was studied. Whole cashew nuts, subjected to low-temperature (LT) and high-temperature (HT) treatments, were used to determine the antioxidant activity of products. Antioxidant activities of cashew nut, kernel, and testa phenolics extracted increased as the roasting temperature increased. The highest activity, as determined by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity, oxygen radical absorbance capacity (ORAC), hydroxyl radical scavenging capacity, Trolox equivalent antioxidant activity (TEAC), and reducing power, was achieved when nuts were roasted at 130 °C for 33 min. Furthermore, roasting increased the total phenolic content (TPC) in both the soluble and bound extracts from whole nut, kernel, and testa but decreased that of the proanthocyanidins (PC) except for the soluble extract of cashew kernels. In addition, cashew testa afforded a higher extract yield, TPC, and PC in both soluble and bound fractions compared to that in whole nuts and kernels. Phenolic acids, namely, syringic (the predominant one), gallic, and p-coumaric acids, were identified. Flavonoids, namely, (+)-catechin, (-)-epicatechin, and epigallocatechin, were also identified, and their contents increased with increasing temperature. The results so obtained suggest that HT-short time (HTST) roasting effectively enhances the antioxidant activity of cashew nuts and testa.     

Kinetic changes in glucosinolate-derived volatiles by heat-treatment and myrosinase activity in nakajimana (Brassica rapa L. cv. nakajimana)

Nakajimana (Brassica rapa L. cv. nakajimana), of the family Brassicaceae, is a traditional vegetable in Japan. Three isothiocyanates and five cyanides in the leaves of nakajimana were identified by gas chromatography (GC) and GC-mass spectrometry (GC-MS), and their kinetic changes using heat-treatment (temperature and time) were investigated. In addition, myrosinase activity of extracts prepared from fresh nakajimana leaf was determined. In crushed heat-treated leaves of nakajimana (70 °C for 30 s), formation of isothiocyanates and myrosinase activity increased, whereas formation of 1-cyano-3,4-epithiobutane and 1-cyano-4,5-epithiopentane decreased. Heat-treatment can significantly alter the content of potentially beneficial compounds in nakajimana, and ingestion of suitable isothiocyanates for human health may be better facilitated by mild boiling.     

Influence of industrial and alternative farming systems on contents of sugars, organic acids, total phenolic content, and the antioxidant activity of red beet (Beta vulgaris L. ssp. vulgaris Rote Kugel)

The contents of sugars, organic acids, total phenolic content, and the antioxidant activity were quantified in the flesh of red beet from conventional (CON), integrated (INT), organic (ORG), biodynamic (BD), and control farming systems using established methods. Significant differences were measured for malic acid, total phenolic content (TPC), and total antioxidant activity, where malic acid content ranged from 2.39 g kg(-1) FW (control) to 1.63 g kg(-1) FW (CON, ORG, and INT). The highest TPC was measured in BD and control samples (0.677 and 0.672 mg GAE g(-1), respectively), and the lowest in CON samples (0.511 mg GAE g(-1)). Antioxidant activity was positively correlated with TPC (r2=0.6187) and ranged from 0.823 μM TE g(-1) FW to 1.270 μM TE g(-1) FW in CON and BD samples, respectively, whereas total sugar content ranged from 21.03 g kg(-1) FW (CON) to 31.58 g kg(-1) FW (BD). The importance of sugars, organic acids, phenols, and antioxidants for human health, as well as for plant resilience and health, gained from this explorative study, is discussed and put into perspective.     

Kaiware Daikon (Raphanus sativus L.) extract: a naturally multipotent chemopreventive agent

Brassica vegetables are attracting major attention as healthy foods because of their content of glucosinolates (GLs) that release the corresponding isothiocyanates (ITCs) upon myrosinase hydrolysis. A number of studies have so far documented the chemopreventive properties of some ITCs. On the other hand, single nutrients detached from the food itself risk being somewhat "reductive", since plants contain several classes of compounds endowed with a polyhedral mechanism of action. Our recent finding that 4-methylthio-3-butenyl isothiocyanate (GRH-ITC) and 4-methylsulfinyl-3-butenyl isothiocyanate (GRE-ITC), released by the GLs purified from Japanese (Kaiware) Daikon (Raphanus sativus L.) seeds and sprouts, had selective cytotoxic/apoptotic activity on three human colon carcinoma cell lines prompted further research on the potential chemopreventive role of a standardized Kaiware Daikon extract (KDE), containing 10.5% w/w GRH and 3.8% w/w GRE, compared to its isolated components. KDE administered in combination with myrosinase at doses corresponding to 50 microM GRH-ITC plus 15 microM GRE-ITC (50 microM KDE-ITC) to three human cancer cell lines (LoVo, HCT-116 and HT-29) significantly reduced cell growth by 94-96% of control in six days (p < 0.05), outperforming pure GRH-ITC or GRE-ITC at the same dose. On the other hand, the same treatment had no significant toxicity on normal human T-lymphocytes. A 50 microM concentration of KDE-ITC had relevant apoptosis induction in all tested cancer cell lines, as confirmed by annexin V assay (e.g., 33% induction in LoVo compared to control, p < 0.05), Bax protein induction (e.g., +20% in HT-29, p < 0.05), and Bcl2 downregulation (e.g.-20% in HT-29, p < 0.05), and induced caspase-1 and PARP-1 activation in all cancer cells as shown by Western blot analysis. Unlike pure GRH or GRH-ITC, KDE also had significant chain-breaking antioxidant activity, retarding the AAPH-initiated autoxidation of methyl linoleate in SDS micelles at concentrations as low as 4.4 ppm (-50% in oxygen consumption rate), as monitored by Clark-type microelectrode oxygen-uptake kinetics, and induced very fast quenching of DPPH. radical in methanol with t(1/2) (s) = (1.47 +/- 0.25) x 10(-2)/[KDE; (g/L)], measured by stopped-flow UV-vis kinetics at 298 K. The potential chemopreventive role of KDE is discussed.     

Antioxidant constituents of radish sprout (Kaiware-daikon), Raphanus sativus L

Methanol extracts of 11 kinds of commonly available vegetables were examined for hydroxyl radical scavenging potency using the bleomycin-Fe method. In this method, the iron ion and bleomycin in water form hydroxyl radicals, and the scavenging activity is monitored by the modified thiobarbituric acid method. All extracts showed scavenging capacity, even though the activity of some of them was lower than that of l-ascorbic acid. Those vegetables were classified into three groups according to their activity, groups showing strong activity, moderate activity, and weak activity, as compared to the activity of l-ascorbic acid at the same concentration. Among them, the methanol extract of radish sprout (Japanese name "kaiware-daikon") exhibited the highest potency (1.8 times as l-ascorbic acid). Then, we investigated the constituents of the methanol extract of radish sprout and the contribution to the overall activity of each compound by examining their activity. As the result, several kinds of sinapinic acid esters and flavonoids were isolated with high radical scavenging potency, which must contribute substantially to the activity.     

秸秆还田方式对土壤质量和萝卜产量及品质的影响

本文以萝卜（品种为枇杷叶）为研究对象，采用田间试验探究了无秸秆还田（CK），秸秆粉碎还田（SC）、过腹还田（SM）、炭化还田（SB）、粉碎+过腹还田（SC+SM）、粉碎+炭化还田（SC+SB）、过腹+炭化还田（SM+SB）对土壤理化性质和萝卜产量及品质的影响。结果表明，6种秸秆还田方式均显著提高了土壤有机质和全氮含量，但对土壤速效钾含量的影响不同，其中SC、SM和SB 3种还田方式不影响土壤速效钾含量，而SC+SM、SC+SB和SM+SB 3种秸秆组合还田方式显著提高了土壤速效钾含量；不同秸秆还田方式对萝卜生物量和叶片叶绿素相对含量（SPAD值）的影响不同，除SC+SM处理萝卜肉质根鲜物质量、叶片SPAD值显著低于对照外，其他秸秆还田处理萝卜肉质根鲜物重量和叶片SPAD值均显著高于对照；秸秆还田处理萝卜品质较对照显著提升，6种秸秆还田处理萝卜维生素C、可溶性糖、总酚、类黄酮含量均显著高于对照，且不同还田方式间无差异；秸秆还田还显著降低了萝卜硝酸盐含量，提高了可溶性蛋白含量，其中SC+SM、SC+SB和SM+SB 3种秸秆组合还田处理表现突出，硝酸盐含量降低36.04%~40.63%，可溶性蛋白含量提升41.70%~56.11%； SC、SM和SB处理次之，硝酸盐含量降低15.15%~22.58%，可溶性蛋白含量提升16.61%~25.11%； 6种秸秆还田方式下萝卜铁离子还原能力（FRAP）显著高于对照，但1，1-二苯基-2-三硝基苯肼自由基清除能力（DPPH）不同，其中SM、SC+SM、SC+SB和SM+SB处理萝卜DPPH值显著高于对照，SC和SB处理萝卜DPPH值与对照接近。综上所述，除SC+SM处理降低了萝卜产量外，SC、SM、SB、SC+SB、SM+SB等5个秸秆还田处理均提高了土壤肥力和作物产量、改善了产品品质，其中SC+SB、SM+SB在降低萝卜硝酸盐含量方面表现尤为突出。考虑到操作的难易程度，常规蔬菜生产中建议采用秸秆粉碎还田、过腹还田、炭化还田3种方式，种植根菜、叶菜等硝酸盐高积累型蔬菜时建议采用粉碎+炭化还田、过腹+炭化还田2种秸秆组合还田模式。     

Fermentation as a bio-process to obtain functional soybean flours

The effect of fermentation on the antioxidant compounds [vitamins C and E, total phenolic compounds (TPC), and reduced glutathione (GSH)], and antioxidant capacity [superoxide anion scavenging activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), inhibition of phosphatidylcholine (PC) peroxidation, and Trolox equivalent antioxidant capacity (TEAC)] of soybean (Glycine max cv. Merit) was studied. Fermentation was carried out in solid state in cracked seeds inoculated with Aspergillus oryzae, Rhizopus oryzae, Bacillus subtilis, and Lactobacillus plantarum and in liquid state either in cracked seeds or milled soybean flours fermented naturally by only the microorganisms present in the seeds or by inoculation with L. plantarum. Vitamin C was not detected in the studied samples. Fermentation caused a decrease in vitamin E activity, except when cracked seed was fermented with A. oryzae, R. oryzae, or B. subtilis that increased 31, 30, and 89%, respectively. Fermentation produced an increase in TPC content and did not affect or reduce the GSH content. Fermentation decreased SOD-like activity drastically, while PRTC increased except when it was carried out naturally in cracked seed. TEAC values rose sharply when soybeans were fermented with B. subtilis. Processed soybean extracts inhibited PC peroxidation in comparison with the control assay. On the basis of the results obtained, the relative contributions of vitamin E, TPC, and GSH to antioxidant capacity were calculated and results showed a very high TPC contribution and a low contribution of GSH and vitamin E activity. Optimum results for functional soybean flours were achieved when fermentation was carried out with B. subtilis inoculum.