Effect of Different Treatment on Anther Callus Formation and Browning in Balsam Pear （Mornordica charantia L.）

Brown callus derived from anther limited the application of anther culture in balsam pear. In order to establish a more perfect regeneration system from anther cultuer, the effects of low temperature pretreatment, 2,4-dichlorophenoxyacetic acid （2,4-D）, vitamin C （Vc） and silver nitrate （AgNO3） on callus induction and browning in anther culture of balsam pear （Momordiea charantia L.） were investigated. The results showed that after pretreatment at 4 ℃ for 1 day, callus induction rate was the highest and browning rate was the lowest. Anthers on MS medium supplemented with 2,4-D 0.5 mg/L formed more and better callus. The medium supplemented with Vc or AgNO3 was advantageous to the induction of callus and reduction of browning. When cultured on medium supplemented with 50 mg/L Vc or 5 mg/L AgNO3, callus induction rate was the highest and browning rate was rather low.

Prevalence of vegetation browning in China’s drylands under climate change

Vegetation greening has long been acknowledged,but recent studies have pointed out that vegetation greening is possibly stalled or even reversed.However,detailed analyses about greening reversal or increased browning of vegetation remain scarce.In this study,we utilized the normalized difference vegetation index(NDVI)as an indicator of vegetation to investigate the trends of vegetation greening and browning(monotonic,interruption,and reversal)through the breaks for the additive season and trend(BFAST)method across China’s drylands from 1982 to 2022.It also reveals the impacts of ecological restoration programs(ERPs)and climate change on these vegetation trends.We find that the vegetation displays an obvious pattern of east-greening and west-browning in China’s drylands.Greening trends mainly exhibits monotonic greening(29.8%)and greening with setback(36.8%),whereas browning shows a greening to browning reversal(19.2%).The increase rate of greening to browning reversal is 0.0342/yr,which is apparently greater than that of greening with setback,0.0078/yr.This research highlights that,under the background of widespread vegetation greening,vegetation browning is pro-gressively increasing due to the effects of climate change.Furthermore,the ERPs have significantly increased vegetation coverage,with the increase rate in 2000-2022 being twice as much as that of 1982-1999 in reveg-etation regions.Vegetation browning in southwestern Qingzang Plateau is primarily driven by adverse climatic factors and anthropogenic disturbances,which offset the efforts of ERPs.

Population genomic analysis reveals key genetic variations and the driving force for embryonic callus induction capability in maize

Genetic transformation has been an effective technology for improving the agronomic traits of maize.However,it is highly reliant on the use of embryonic callus(EC)and shows a serious genotype dependence.In this study,we performed genomic sequencing for 80 core maize germplasms and constructed a high-density genomic variation map using our newly developed pipeline(MQ2Gpipe).Based on the induction rate of EC(REC),these inbred lines were categorized into three subpopulations.The low-REC germplasms displayed more abundant genetic diversity than the high-REC germplasms.By integrating a genome-wide selective signature screen and region-based association analysis,we revealed 95.23 Mb of selective regions and 43 REC-associated variants.These variants had phenotypic variance explained values ranging between 21.46 and 49.46%.In total,103 candidate genes were identified within the linkage disequilibrium regions of these REC-associated loci.These genes mainly participate in regulation of the cell cycle,regulation of cytokinesis,and other functions,among which MYB15 and EMB2745 were located within the previously reported QTL for EC induction.Numerous leaf area-associated variants with large effects were closely linked to several REC-related loci,implying a potential synergistic selection of REC and leaf size during modern maize breeding.

Anticancer Activity of Rice Callus Suspension Cultures from Aromatic Varieties and Metabolites Regulated in Treated Cancer Cell Lines

Tissue culture techniques were used to produce large amounts of bioactive compounds with medicinal potential, overcoming space and time constraints for cancer prevention. Rice callus suspension cultures(RCSC) and seed extracts prepared from aromatic rice varieties were used to evaluate the cytotoxic impact on human colon and lung cancer cell lines, as well as a normal control cell line, using Taxol as a positive control. RCSC and seed extracts from two Indian aromatic rice varieties were applied at different concentrations to treat the cancer cell lines and normal lung fibroblasts over varying time intervals. Apoptosis was assessed in 1:5 dilutions of the A549 and HT-29 cell lines treated with RCSC for 72 h, using propidium iodide staining and flow cytometry. RCSC showed a more potent cytotoxic effect than seed extracts with minimal effect on the normal cell line, in contrast to Taxol. Confocal microscopy and flow cytometry further confirmed the apoptotic effect of RCSC. Gas chromatography-mass spectrometry-based metabolic profiling identified metabolites involved in cytotoxicity and highlighted altered pathways. RCSC is proposed as an alternative source for the development of novel anticancer drugs with reduced side effects.

Transcriptome and metabolome analysis of preharvest internal browning in Nane plum fruit caused by high temperature

The preharvest internal browning of Nane plum fruit,with no visible effects on the appearance of the fruit,has become a serious problem in recent years in its production area in Guangdong Province,China.This study investigated the effects of environmental factors,including temperature,on Nane plum internal browning.Plum orchards at different elevations with different incidences of internal browning were selected.Using fruits with different internal browning incidence levels,the internal browning mechanism was analyzed with transcriptome and metabolome analyses.The results revealed decreased internal browning at high altitudes.Shading treatment significantly reduced internal browning,whereas bagging and insect-proof net-covering treatments significantly increased internal browning.Because bagging and net coverings increase the local ambient temperature,the findings suggest that high temperature is an important factor influencing the internal browning of Nane plum.The metabolome experiments showed that with increased internal browning,the levels of phenolic hydroxyls such as catechol increased,with simultaneous increases in hydrogen peroxide content and oxidase activity.It can be speculated that the oxidation of phenolic hydroxyl substances is the main cause of the preharvest browning of Nane plum.Transcriptome analysis revealed the increased expression of calcium signaling-related and downstream effector genes and indicated an important role of calcium in internal browning,possibly due to its increased content in the fruit.Further,with increasingly serious internal browning,genes related to photosynthesis were down-regulated,while genes related to senescence were up-regulated,thus suggesting the up-regulation of the process of cell senescence during internal browning.In conclusion,heat stress should be eliminated to reduce preharvest internal browning in Nane plum.

Resistance index and browning mechanism of apple peel under high temperature stress

Apples are one of the most important economic crops worldwide.Because of global warming and an aggravation of environmental,abnormally high temperatures occur frequently in fruit-growing season and seriously affect normal fruit growth and reduce fruit quality and yield.We took five-year-old Ruixue’(Qinfu 1×Pink Lady;CNA20151469.1) fruits as test materials,and the ambient temperature during fruit development was monitored.The results showed that during the fruit-growing season,especially during the rapid growth stage (July to August),the maximum daily temperature exceeded 30℃ and lasted for more than 40 days.To determine the effects of high temperature stress on the apple fruit resistance,we treated expanding,veraison,and maturity-period fruits at different temperatures.It was found that the fruits of the expanding period showed strong resistance to high temperature stress,whereas during veraison and maturity,fruit resistance to high temperature stress decreased,and the fruit peel browning phenotype appeared.Meanwhile,the content of malonaldehyde (MDA),hydrogen peroxide (H_(2)O_(2)),and superoxide anion (O._(2)^(-)) in the peel gradually increased with increasing temperature.The content of total phenols,flavanol,and flavonoids in the peel decreased substantially at 45℃.Moreover,it was found that polyphenol oxidase gene (MdPPO1) was most sensitive to high temperature stress in apple.Furthermore,transient and stable MdPPO1 overexpression significantly promoted peel browning.The transgenic materials were more sensitive to high temperatures,and browning was more severe compared to non-genetically modified organism (WT).Stable MdPPO1 knockout calli obtained via clustered regularly interspersed short palindromic repeats (CRISPR/Cas9) gene knockout technology reduced the browning phenotype,and the resultant fruits were not sensitive to the effects of high temperature stress.Thus,MdPPO1 expression may be a key factor of high temperature-related changes observed in the browning phenotype that provides a scientific theoretical basis for the selection of high temperature-resistant varieties and apple cultivation and management in the future.

Ekphrasis in Robert Browning’s “Fra Lippo Lippi”

As an ekphrastic poem re-created from the painting Coronation of the Virgin,“Fra Lippo Lippi”lacks adequate scrutiny from the perspective of interart poetics.With regards to the core concept of ekphrasis,this paper makes an in-depth analysis of the features and functions of ekphrasis in the poem.The former lie in the poet’s identification with the painter,and his imaginative supplementation in“the verbal representation of visual representation”.The latter are embodied in the revelation of the invisible in the painting,namely,the desires and value of human beings,as well as the power of beauty and truth.Browning enriches the poem with both philosophical and artistic contemplation,thus strongly conveying the thematic implications of“know thyself”and the persistent pursuit of beauty and truth.It is hoped that this study can shed some light on the studies of Robert Browning and the poem itself,and contribute to the development of the interart poetics.

A Preliminary Study on Callus Induction and Proliferation of Cylocarya paliurus

[Objective] To carry out preliminary study on callus induction and prolifer- ation of Cylocarya pafiurus. [Method] The leaf and stem segments of Cylocarya paliurus as explants were cultured on MS, WPM and improved DKW3 mediums, added 6-BA and IBA with different concentration ratios to explore the optimum medium for callus induction and proliferation. [Result] It was found that MS ＋ 4.0 mg/L 6-BA ＋ 2.0 mg/L IBA was the optimum medium for inducing stem callus of Cylocarya pali- urus, MS ＋ 2.0 mg/L 6-BA ＋ 0.5 mg/L IBA was the optimum medium for inducing leaf explants callus of Cylocarya paliurus, and MS ＋ 1.0 mg/L 6-BA ＋ 1.0 mg/L IBA was the optimum medium for callus proliferation. 200 mg/L Vitamin C was best to inhibit Cylocarya paliurus callus from browning with the browning rate of only 5.71%. [Conclusion] This study provided some theoretical basis for the establishment of tis- sue culture and rapid propagation system and the development of molecular breed- ing study of Cylocarya paliurus.

Effect of Different Disinfection Methods on Calluses of Amorphophallus albus P.Y.Liu&J.F.Chen

This study aimed to examine the effect of different disinfection methods on the callus of Amorphophallus albus in the tissue culture and rapid propagation system.The calluses induced from buds of A.albus were disinfected with benzalkonium bromide,mercury bichloride,sodium hypochlorite,and ethanol,respectively,for different time periods.The growth of calluses was compared between different treatments and observed in detail by dissection.The results showed that the disinfection with 0.1% mercury bichloride for 5 min and 75% ethanol for 5 s had mild effect on the callus,with the lowest browning rates of 16.32% and 15.46% and the lowest vitrification rates of 3.24% and 9.72%,respectively,after 20 d.The disinfection with benzalkonium bromide and sodium hypochlorite had strong effect on the callus cells,with the lowest browning rate of 50.69% and 25.12% and the lowest vitrification rates of 28.47% and 28.43%,respectively.The cytological observation result showed that the calluses disinfected with ethanol and mercury bichloride showcased compact structures,while those disinfected with sodium hypochlorite and benzalkonium bromide exhibited loose structures.To sum up,it is recommended that 75% ethanol or 0.1% mercury bichloride can be used for disinfection in the tissue culture and rapid propagation of A.albus,while the use of benzalkonium bromide and sodium hypochlorite should be avoided as much as possible.

Optimization of Inducing Conditions for Loose Callus of Pinellia ternata

[Objective] The research aimed to establish the cell suspension culture system of Pinellia ternate.[Method] The petiole of Pinellia ternate was as the explant.The orthogonal test was used to study the influences of four plant growth substances（2,4-D,NAA,picloram and KT）and their mixture ratios on the formation of loose callus.[Result] The induction effect of 2,4-D and picloram on the petiole callus of Pinellia ternate was the most significant.Then,the second ones were KT and NAA.The optimal medium which induced Pinellia ternate petiole to form the loose callus was MS＋0.5 mg/L of 2,4-D＋1.0 mg/L of NAA＋1.0 mg/L of picloram＋1.5 mg/L of KT.[Conclusion] The research laid the foundation for extracting the active ingredient from the cell suspension of Pinellia ternate and producing the artificial seed.

Sugarcane Callus Culture and Cytological Characteristics

In order to obtain sugarcane embryogenic calli for transgenosis, the effects of different soaking time of explants, medium, culture conditions and sampling time on sugarcane callus culture were investigated. The results showed that soaking ex- plants with MS liquid medium for 20 min could significantly reduce the browning rate; MS＋3.0 mg/L 2.4-D was appropriate for induction and proliferation of embryo- genic calli; dark condition was suitable for the culture of sugarcane calli; calli differ- entiated from materials collected in August exhibited low browning rate, high callus induction rate and high embryogenic callus induction rate. Calli with different pheno- types were stained, prepared into slides and observed under a microscope to ana- lyze the cytological characteristics. The resultsshowed that milky-white granular em- bryogenic calli had closely arranged cells with large nucleus and exhibited strong differentiation capacity, which were appropriate receptor materials for genetic trans- formation of sugarcane.

Study on the Browning in Cell Suspension Culture of Taxus cuspidata

[Objective] This study aimed to investigate the browning of T. cuspidata cells in suspension culture and provide the guidance for the cell suspension culture of T. cuspidata. [Method] T. cuspidata callus was used as experimental materials, to explore the effect of different medium, N/P ratio, pH, shaking speed, illumination time and light intensity and other factors on browning of T. cuspidata cells in suspension culture. [Result] Non-browning callus was transferred to 2MB5 medium （pH 7.0） for illumination culture at 22℃ under light intensity of 1 500 lx with shaking speed of 90 r/min for 24 h. Results showed that the cell browning was significantly inhibited. [Conclusion] This study laid the foundation for cell suspension culture of T. cuspidata and had important significance to the large-scale industrial production of paclitaxel.

Chemical profiling of bioactive compounds in the methanolic extract of wild leaf and callus of Vitex negundo using gas chromatographymass spectrometry

BACKGROUND The investigation of plant-based therapeutic agents in medicinal plants has revealed their presence in the extracts and provides the vision to formulate novel techniques for drug therapy.Vitex negundo(V.negundo),a perennial herb belonging to the Varbanaceae family,is extensively used in conventional medication.AIM To determine the existence of therapeutic components in leaf and callus extracts from wild V.negundo plants using gas chromatography-mass spectrometry(GCMS).METHODS In this study,we conducted GC-MS on wild plant leaf extracts and correlated the presence of constituents with those in callus extracts.Various growth regulators such as 6-benzylaminopurine(BAP),2,4-dichlorophenoxyacetic acid(2,4-D),α-naphthylacetic acid(NAA),and di-phenylurea(DPU)were added to plant leaves and in-vitro callus and grown on MS medium.RESULTS The results clearly indicated that the addition of BAP(2.0 mg/L),2,4-D(0.2 mg/mL),DPU(2.0 mg/L)and 2,4-D(0.2 mg/mL)in MS medium resulted in rapid callus development.The plant profile of Vitex extracts by GC-MS analysis showed that 24,10,and 14 bioactive constituents were detected in the methanolic extract of leaf,green callus and the methanolic extract of white loose callus,respectively.CONCLUSION Octadecadienoic acid,hexadecanoic acid and methyl ester were the major constituents in the leaf and callus methanolic extract.Octadecadienoic acid was the most common constituent in all samples.The maximum concentration of octadecadienoic acid in leaves,green callus and white loose callus was 21.93%,47.79%and 40.38%,respectively.These findings demonstrate that the concentration of octadecadienoic acid doubles in-vitro compared to in-vivo.In addition to octadecadienoic acid;butyric acid,benzene,1-methoxy-4-(1-propenyl),dospan,tridecanedialdehyde,methylcyclohexenylbutanol,chlorpyrifos,n-secondary terpene diester,anflunine and other important active compounds were also detected.All these components were only available in callus formed in-vitro.This study showed that the callus contained additional botanical characteristics compared with wild plants.Due to the presence of numerous bioactive compounds,the medical use of Vitex for various diseases has been accepted and the plant is considered an important source of therapeutics for research and development.

Physiological Changes during the Process of Pericarp Browning in the Postharvest Litchi

[Objective] Pericarp browning in the postharvest litchi significantly reduced its commercial value and limited the expanding of litchi markets. Physiological changes during the process of pericarp browning were determined in order to identify the underlying mechanisms. [Method] Matured Feizixiao fruits were stored at 25 ℃ and 70%±5% relative humidity. The physiological changes happened in pericarp during storage were tested at an 8-hour interval. [Result] The fruit of Feizixiao (Litchi chinensis Sonn. cv Feizixiao) turned completely brown within 72 h after being harvested under the experimental conditions. Sharp increase of the browning index occurred from 48 to 64 hours after harvest (HAH). With the browning of pericarp,water content of the whole fruit and pericarp decreased continuingly. In contrast,there were no significant changes in the water content of pulp during the same period. MDA content,pH value and relative leakage rate of pericarp were increased during storage. Most of pigment contents including anthocyanin,flavonoid,phenols,chlorophyll a and total chlorophyll decreased. POD activity was initially increased in 32 HAH and then decreased afterwards. PPO activity was decreased continuously,while the activities of catalase and superoxide dismutase exhibited the pattern of 'increasing-decreasing-increasing' as the storage time progressed. Correlation,stepwise regression and path analyses showed that water loss of pericarp was the major factor of pericarp browning. Principal and cluster analyses showed that there were two stages of pericarp browning during the course of litchi storage. [Conclusion] Water status of pericarp was the most important factor affecting pericarp browning. The pericarp browning happened by stages,which was mainly determined by the water loss of pericarp.

A Comparative Study on Detection of the Expression of Alternative Oxidase in Tobacco Callus with the Monoclonal Antibody Against Alternative Oxidase and Antibody Against-Synthetic Polypeptide Antibody

从经过不同温度处理的烟草 (NicotianarusticaL .)愈伤组织中提取并纯化线粒体蛋白 ,分别与交替氧化酶的单克隆抗体和抗合成多肽抗体进行免疫杂交。结果表明 :交替氧化酶的含量随温度的下降而显著上升 ;单克隆抗体的特异性较高于抗合成多肽抗体 ,但后者与交替氧化酶同样有良好的亲和性。因此 ,用合成多肽方法制备的抗体可以用于交替氧化酶的研究中。

Effect of Sound Stimulation on the Lipid Physical States and Metabolism of Plasma Membrane from Chrysanthemum Callus

Plasma membrane vesicles of chrysanthemum ( Dendranthema morifolium (Ramat.) Tzvel.) callus was purified by sucrose gradient centrifugation to investigate the influence of sound stimulation on the lipid physical states and metabolism of plasma membrane. The results showed that sound stimulation decreased the content of phosphodiesters and the fluorescent intensity of DPH, but increased the light scattering value of the membrane, the fluorescent intensity of MC540 and the content of phosphomonoesters, indicating that the vesicles got looser, the charge density and hydrophobicity of membrane surface decreased under sound stimulation of some strength and frequency. However, the membrane fluidity increased under the condition. Meanwhile, the anabolism of membrane lipid increased and the catabolism decreased. It can be seen that the physical state and metabolism of membrane lipid is sensitive to sound stimulation.

Study on Cu Resistance of Ricinus communis L. Callus

[Objective] The article studies the growth and Cu absorption of Ricinus communis L. callus under Cu stress. [Method] CuSO4.5H20 solutions with different Cu concentrations were added to callus subculture medium; callus was inoculated and Cu resistance index of callus was worked out. Cu content in callus was deter- mined with the method of Varian AA240FS. [Result] With the Cu concentration at 60 mg/L, the growth of callus was inhibited, its Cu resistance index was only 33.87%. With the Cu concentration at 40 mg/L, callus was faint yellow in color, and grew rapidly with its Cu resistance index at 61.29%. Such high level resistance could remain the same after six week after continuous subculture. In the 4t week of culture, Cu resistance index in treatments with Cu concentrations at 10, 20, 30, 40 mg/L was higher than that in the 3rd week, and the content of Cu in callus of the treatments was 0.33, 0.54, 1.16, 1.40 mg/g respectively. [Conclusion] Cu con- centration at 40 mg/L in culture medium can be the threshold for selecting Cu re- sistance R. communis callus.

On the Dramatic Features in Robert Browning's Poems of Dramatic Monologue

This paper explored the three dramatic features intrinsic in Robert Browning's poems of dramatic monologue, namely, the employment of first-person point of view, the use of irony, and the intricate psychological exploration and vivid characterization. Such features are crucial in the successful presentation of various human nature and experience with profundity, making his poems an enduring classic in the English literature.

Insecticidal Activities of Extracts from Brucea javanica (L.) Merr. Callus

[Objective] The research aimed to lay a foundation for the screening of cell lines producing secondary metabolites of Brucea javanica(L.)Merr.[Method] The insecticidal activities of the extracts from branch and 3 different types of calluses of Brucea javanica(L.)Merr.was detected through methods of leaf disc and potted seedlings against the diamond back moth.[Result] Extracts from four kinds of Brucea javanica(L.)Merr.tissues assumed both the activities of antifeedant and oviposition deterrency against the diamond back moth.Antifeedant effect of extracts was in turn the callus C< callus B< callus A< branches.Oviposition deterrency activity of the extracts was in turn the callus A> branch > callus B>callus C.The insecticidal activities of callus A and B were higher than that of the callus C.[Conclusion] The results show that insecticidal activity of callus and its growth rate is inversely proportional.

Optimization of Callus Induction Medium for Schisandga chinensis Baill via Uniform Design

[Objective]This study was to optimize callus induction medium for Schisandga chinensis Baill.[Method]Using callus induction rate as an indicator,uniform design was employed to optimize hormone combination at poly-factors and poly-levels for callus induction from Schisandga chinensis Baill.[Results]Optimal hormone combinations depended on different explants：optimum medium for both tender leaf and petiole was MS ＋3 mg/L 6-BA,for stem segment was MS ＋3 mg/L 6-BA ＋0.6 mg/L NAA.[Conclusion]Uniform design is a time-saving and convenient method for the optimum medium for callus and yields a higher callus induction rate.