caa a22 4500 606208399 CHVBK 20210128101022.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00425-015-2345-x doi (NATIONALLICENCE)springer-10.1007/s00425-015-2345-x Biotic stress-induced expression of mulberry cystatins and identification of cystatin exhibiting stability to silkworm gut proteinases [Elektronische Daten] [Jiubo Liang, Yupeng Wang, Guangyu Ding, Wensheng Li, Guangwei Yang, Ningjia He] Main conclusion : Biotic stresses induce the expression of mulberry cystatins. MaCPI-4 protein is stable in silkworm digestive fluid and accumulates in gut food debris and frass. Plant cystatins are considered to be involved in defense responses to insect herbivores though little is known about how cystatins from the natural host respond to a specialist herbivory and the following postingestive interaction is also poorly understood. Here, we studied the biotic stress-mediated inductions of cystatins from mulberry tree, and examined the stability of mulberry cystatin proteins in the gut of silkworm, Bombyx mori, a specialist insect feeding on mulberry leaf. First, we cloned and characterized six cystatin genes from a mulberry cultivar, Morus atropurpurea Roxb., named as MaCPI-1 to MaCPI-6. The recombinant MaCPI-1, MaCPI-3 and MaCPI-4 proteins, which showed inhibitory effects against papain in vitro, were produced. Silkworm herbivory as well as methyl jasmonate (MeJA) treatment induced the expression of five mulberry cystatin genes, and the highest inductions were observed from MaCPI-1 and MaCPI-6. Mechanical wounding led to the inductions of four cystatin genes. The differential induction occurred in MaCPI-2. The induced protein changes were detected from three mulberry cystatins comprising MaCPI-1, MaCPI-3 and MaCPI-4. In vivo and in vitro assays showed that MaCPI-1 and MaCPI-3 proteins were susceptible to silkworm digestive fluid and MaCPI-4 had an antidigestive stability, and was detected in silkworm gut and frass. Collectively, our data indicated that biotic stresses resulted in the transcriptional inductions and protein changes of mulberry cystatins (MaCPIs), and identified MaCPI-4 with stability in the gut of its specialist herbivore. Springer-Verlag Berlin Heidelberg, 2015 Antidigestive stability nationallicence Cystatin nationallicence Induced gene expression nationallicence Insect herbivory nationallicence Mulberry nationallicence CPI : Cysteine proteinase inhibitor nationallicence MeJA : Methyl jasmonate nationallicence PhyCys : Phytocystatins nationallicence PMSF : Phenylmethylsulfonyl fluoride nationallicence Liang Jiubo State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut Wang Yupeng State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut Ding Guangyu State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut Li Wensheng State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut Yang Guangwei State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut He Ningjia State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut Planta Springer Berlin Heidelberg 242/5(2015-11-01), 1139-1151 0032-0935 242:5<1139 2015 242 425 https://doi.org/10.1007/s00425-015-2345-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00425-015-2345-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Liang Jiubo State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut NATIONALLICENCE 700 1- Wang Yupeng State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut NATIONALLICENCE 700 1- Ding Guangyu State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut NATIONALLICENCE 700 1- Li Wensheng State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut NATIONALLICENCE 700 1- Yang Guangwei State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut NATIONALLICENCE 700 1- He Ningjia State Key Laboratory of Silkworm Genome Biology, Southwest University, 400715, Chongqing, China aut NATIONALLICENCE 773 0- Planta Springer Berlin Heidelberg 242/5(2015-11-01), 1139-1151 0032-0935 242:5<1139 2015 242 425