Silage Cassia alata (CA) can alleviate feed shortage in some areas to a certain extent and reduce feed costs. The present research evaluated the effect of cellulase (CE) and Lactiplantibacillus plantarum (LP) on the fermentation parameters, nutrients, and bacterial community of CA silage. Chopped CA was ensiled with three different treatments, namely, no inoculant (CK), CE, and LP, and the indexes were determined on the 2nd, 6th, 14th, and 30th days of silage fermentation. The fermentation parameters indicate that the pH value of the three groups decreased and then increased with the ensilage process, and the lowest value was observed on the 14th day. The CK and LP groups attained the highest value on the 30th day, while the CE group attained the highest value on the 2nd day. Additionally, the pH value and NH3-N content were significantly lower (P < 0.05) in the CE and LP groups than in the CK group. In terms of nutrients, crude protein (CP) contents significantly increased (P < 0.05) in the CE and LP groups on the 30th day. The neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents of the CE group were significantly and negatively associated with fermentation time, and the water-soluble carbohydrate (WSC) contents of the three groups were significantly lower during ensiling. In comparison with the CK group, the NDF and ADF contents were significantly reduced (P < 0.05), and the WSC content increased (P < 0.05) in the CE group on day 30. Sequencing analysis of bacterial communities showed that Lactobacillus became the most dominant genus in the ensilage process. Moreover, both CE and LP groups increased the abundance of Lactobacillus and decreased that of Klebsiella, Weissella, and Acetobacter in comparison to the CK group, in which LP had a better effect. CE and LP could further improve the silage quality of CA, and LP had a more significant effect in reconstructing the bacterial community in the silage environment.

The Cassia (Leguminosae) genus has attracted a lot of attention as a prolific source of alkaloids and chromones with diverse structures and biological properties. The aim of this study is to screen the antiviral compounds from Cassia alata. The extract of the stem bark of this plant was separated using silica gel, MCI, ODS C18, and Sephadex LH-20 column chromatography, as well as semi-preparative HPLC. As a result, three new indole alkaloids, alataindoleins A-C (1-3); one new chromone, alatachromone A (4); and a new dimeric chromone-indole alkaloid, alataindolein D (5) were isolated. Their structures were determined by means of HRESIMS and extensive 1D and 2D NMR spectroscopic studies. Interestingly, alataindolein D (5) represents a new type of dimeric alkaloid with an unusual N-2-C-16\' linkage, which is biogenetically derived from a chromone and an indole alkaloid via an intermolecular nucleophilic substitution reaction. Compounds 1-5 were tested for their anti-tobacco mosaic virus (TMV) and anti-rotavirus activities, and the results showed that compounds 2-4 showed high anti-TMV activities with inhibition rates of 44.4%, 66.5%, and 52.3%, respectively. These rates were higher than those of the positive control (with inhibition rate of 32.8%). Compounds 1 and 5 also showed potential anti-TMV activities with inhibition rates of 26.5% and 31.8%, respectively. In addition, compounds 1-5 exhibited potential anti-rotavirus activities with therapeutic index (TI) values in the range of 9.75~15.3. The successful isolation and structure identification of the above new compounds provided materials for the screening of antivirus drugs, and contributed to the development and utilization of C. alata.

Two structurally unique polyphenols, alatains A (1) and B (2), were isolated from the bark of Cassia alata. Their structures were elucidated on the basis of spectroscopic analysis. Compounds 1 and 2 represent a new type of hetero-dimeric polyphenols with a C-14-C-5\' linkage, biogenetically formed by an unusual intermolecular oxidative phenol-coupling reaction between a chromone unit and an isocoumarin moiety. Moreover, compounds 1 and 2 showed significant anti-tobacco mosaic virus (anti-TMV) inhibition IC50 values of 18.8 and 11.4 μM, respectively. Alatains A and B also exhibited promising protective effects on TMV infection of the host plants (Nicotiana tabacum) with the inhibition rates of 54.6% and 69.7% at the concentration of 20 μM, respectively. The results provided a new structural template for potential anti-TMV agent discovery.