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SUMMARY: The potato's most devastating disease is late blight, which is caused by Phytophthora infestans. Whereas various resistance (R) genes are known, most are typically defeated by this fast‐evolving oomycete pathogen. However, the broad‐spectrum and durable R8 is a vital gene resource for potato resistance breeding. To support an educated deployment of R8, we embarked on a study on the corresponding avirulence gene Avr8. We overexpressed Avr8 by transient and stable transformation, and found that Avr8 promotes colonization of P. infestans in Nicotiana benthamiana and potato, respectively. A yeast‐two‐hybrid (Y2H) screen showed that AVR8 interacts with a desumoylating isopeptidase (StDeSI2) of potato. We overexpressed DeSI2 and found that DeSI2 positively regulates resistance to P. infestans, while silencing StDeSI2 downregulated the expression of a set of defense‐related genes. By using a specific proteasome inhibitor, we found that AVR8 destabilized StDeSI2 through the 26S proteasome and attenuated early PTI responses. Altogether, these results indicate that AVR8 manipulates desumoylation, which is a new strategy that adds to the plethora of mechanisms that Phytophthora exploits to modulate host immunity, and StDeSI2 provides a new target for durable resistance breeding against P. infestans in potato. Significance Statement: StDeSI2, identified as a key host target of AVR8 in potato, plays a positive role in regulating late blight resistance. The present research enriches our knowledge about a new strategy that P. infestans uses to control host defense, and StDeSI2 promises to provide a new target for potato breeding against late blight. [ABSTRACT FROM AUTHOR]

The Potato virus Y (PVY) is responsible for huge economic losses for the potato industry worldwide and is the fifth most consequential plant virus globally. The main strategies for virus control are to limit aphid vectors, produce virus-free seed potatoes, and breed virus-resistant varieties. The breeding of PVY-resistant varieties is the safest and most effective method in terms of cost and environmental protection. Rychc, a gene that confers extreme resistance to PVY, is from S. chacoense, which is a wild diploid potato species that is widely used in many PVY-resistant breeding projects. In this study, Rychc was fine mapped and successfully cloned from S. chacoense accession 40-3. We demonstrated that Rychc encodes a TIR-NLR protein by stably transforming a diploid susceptible cultivar named AC142 and a tetraploid potato variety named E3. The Rychc conferred extreme resistance to PVYO, PVYN:O and PVYNTN in both of the genotypes. To investigate the genetic events occurring during the evolution of the Rychc locus, we sequenced 160 Rychc homologs from 13 S. chacoense genotypes. Based on the pattern of sequence identities, 160 Rychc homologs were divided into 11 families. In Family 11 including Rychc, we found evidence for Type I evolutionary patterns with frequent sequence exchanges, obscured orthologous relationships and high non-synonymous to synonymous substitutions (Ka/Ks), which is consistent with rapid diversification and positive selection in response to rapid changes in the PVY genomes. Furthermore, a functional marker named MG64-17 was developed in this study that indicates the phenotype with 100% accuracy and, therefore, can be used for marker-assisted selection in breeding programs that use S. chacoense as a breeding resource. [ABSTRACT FROM AUTHOR]

HSP40 (also known as DnaJ), HSP70, and HSP90 are major heat shock protein (HSP) families that play critical roles in plant growth and development and stress adaption. Recently, several members of the three HSP families were reported to be widely involved in the plant host-virus interactions. However, their global expression profiles and core members recruited by viruses are largely unknown. In this study, a total of 89 StDnaJs were identified from a genome-wide survey, and their classification, phylogenetic relationships, chromosomal locations, and gene duplication events were further analyzed. Together with 20 StHSP70s and 7 StHSP90s previously identified in the potato genome, the global expression patterns of the members in 3 HSP families were investigated in 2 potato cultivars during Potato virus Y (PVY) infection using RNA-seq data. Of them, 16 genes (including 8 StDnaJs, 6 StHSP70s, and 2 StHSP90s) were significantly up- or downregulated. Further analysis using qRT-PCR demonstrated that 7 of the 16 genes (StDnaJ06, StDnaJ17, StDnaJ21, StDnaJ63, StHSP70-6, StHSP70-19, and StHSP90.5) were remarkably upregulated in the potato cultivar 'Eshu 3' after PVY infection, implying their potential roles in the potato-PVY compatible interaction. Subsequent virus-induced gene silencing (VIGS) assays showed that silencing of the homologous genes of StDnaJ17, StDnaJ21, StHSP70-6, and StHSP90.5 in Nicotiana. benthamiana plants dramatically reduced the accumulation of PVY, which indicated the four genes may function as susceptibility factors in PVY infection. This study provides candidate genes for exploring the mechanism of potato-PVY compatible interaction and benefits breeding work aiming to produce new cultivars with the ability to grow healthily under PVY infection. [ABSTRACT FROM AUTHOR]

Ralstonia solanacearum , which causes bacterial wilt in a broad range of plants, is considered a "species complex" due to its significant genetic diversity. Recently, we have isolated a new R. solanacearum strain HA4-1 from Hong'an county in Hubei province of China and identified it being phylotype I, sequevar 14M (phylotype I-14M). Interestingly, we found that it can cause various disease symptoms among different potato genotypes and display different pathogenic behavior compared to a phylogenetically related strain, GMI1000. To dissect the pathogenic mechanisms of HA4-1, we sequenced its whole genome by combined sequencing technologies including Illumina HiSeq2000, PacBio RS II, and BAC-end sequencing. Genome assembly results revealed the presence of a conventional chromosome, a megaplasmid as well as a 143 kb plasmid in HA4-1. Comparative genome analysis between HA4-1 and GMI1000 shows high conservation of the general virulence factors such as secretion systems, motility, exopolysaccharides (EPS), and key regulatory factors, but significant variation in the repertoire and structure of type III effectors, which could be the determinants of their differential pathogenesis in certain potato species or genotypes. We have identified two novel type III effectors that were probably acquired through horizontal gene transfer (HGT). These novel R. solanacearum effectors display homology to several YopJ and XopAC family members. We named them as RipBR and RipBS. Notably, the copy of RipBR on the plasmid is a pseudogene, while the other on the megaplasmid is normal. For RipBS, there are three copies located in the megaplasmid and plasmid, respectively. Our results have not only enriched the genome information on R. solanacearum species complex by sequencing the first sequevar 14M strain and the largest plasmid reported in R. solanacearum to date but also revealed the variation in the repertoire of type III effectors. This will greatly contribute to the future studies on the pathogenic evolution, host adaptation, and interaction between R. solanacearum and potato. [ABSTRACT FROM AUTHOR]

Summary: Both Solanum tuberosum and Ralstonia solanacearum phylotype IIB originated in South America and share a long‐term co‐evolutionary history. However, our knowledge of potato bacterial wilt pathogenesis is scarce as a result of the technical difficulties of potato plant manipulation. Thus, we established a multiple screening system (virulence screen of effector mutants in potato, growth inhibition of yeast and transient expression in Nicotiana benthamiana) of core type III effectors (T3Es) of a major potato pathovar of phylotype IIB, to provide more research perspectives and biological tools. Using this system, we identified four effectors contributing to virulence during potato infection, with two exhibiting multiple phenotypes in two other systems, including RipAB. Further study showed that RipAB is an unknown protein with a nuclear localization signal (NLS). Furthermore, we generated a ripAB complementation strain and transgenic ripAB‐expressing potato plants, and subsequent virulence assays confirmed that R. solanacearum requires RipAB for full virulence. Compared with wild‐type potato, transcriptomic analysis of transgenic ripAB‐expressing potato plants showed a significant down‐regulation of Ca2+ signalling‐related genes in the enriched Plant–Pathogen Interaction (PPI) gene ontology (GO) term. We further verified that, during infection, RipAB is required for the down‐regulation of four Ca2+ sensors, Stcml5, Stcml23, Stcml‐cast and Stcdpk2, and a Ca2+ transporter, Stcngc1. Further evidence showed that the immune‐associated reactive oxygen species (ROS) burst is attenuated in ripAB transgenic potato plants. In conclusion, a systematic screen of conserved R. solanacearum effectors revealed an important role for RipAB, which interferes with Ca2+‐dependent gene expression to promote disease development in potato. [ABSTRACT FROM AUTHOR]

Cold-induced sweetening (CIS) caused by reducing sugar (RS) accumulation during storage in low temperature in potato tubers is a critical factor influencing the quality of fried potato products. The reconditioning (REC) by arising storage temperature is a common measure to lower down RS. However, both CIS and REC are genotypedependent and the genetic basis remains uncertain. In the present study, with a diploid potato population with broad genetic background, four reproducible QTL of CIS and two of REC were resolved on chromosomes 5, 6, and 7 of the CIS-sensitive parent and chromosomes 5 and 11 of the CIS-resistant parent, respectively, implying that these two traits may be genetically independent. This hypothesis was also supported by the colocalization of two functional genes, a starch synthesis gene AGPS2 mapped in QTL CIS_E_07-1 and a starch hydrolysis gene GWD colocated with QTL REC_B_05-1. The cumulative effects of identified QTL were proved to contribute largely and stably to CIS and REC and confirmed with a natural population composed of a range of cultivars and breeding lines. The present research identified reproducible QTL for CIS and REC of potato in diverse conditions and elucidated for the first time their cumulative genetic effects, which provides theoretical bases and applicable means for tuber quality improvement. [ABSTRACT FROM AUTHOR]

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is an important enzyme that functions in producing energy and supplying intermediates for cellular metabolism. Recent researches indicate that GAPDHs have multiple functions beside glycolysis. However, little information is available for functions of GAPDHs in potato. Here, we identified 4 putative cytosolic GAPDH genes in potato genome and demonstrated that the StGAPC1, StGAPC2, and StGAPC3, which are constitutively expressed in potato tissues and cold inducible in tubers, encode active cytosolic GAPDHs. Cosuppression of these 3 GAPC genes resulted in low tuber GAPDH activity, consequently the accumulation of reducing sugars in cold stored tubers by altering the tuber metabolite pool sizes favoring the sucrose pathway. Furthermore, GAPCs-silenced tubers exhibited a loss of apical dominance dependent on cell death of tuber apical bud meristem (TAB-meristem). It was also confirmed that StGAPC1, StGAPC2, and StGAPC3 interacted with the autophagy-related protein 3 (ATG3), implying that the occurrence of cell death in TAB-meristem could be induced by ATG3 associated events. Collectively, the present research evidences first that the GAPC genes play crucial roles in diverse physiological and developmental processes in potato tubers. [ABSTRACT FROM AUTHOR]

Potato cold-induced sweetening ( CIS) is critical for the postharvest quality of potato tubers. Starch degradation is considered to be one of the key pathways in the CIS process. However, the functions of the genes that encode enzymes related to starch degradation in CIS and the activity regulation of these enzymes have received less attention. A potato amylase inhibitor gene known as Sb AI was cloned from the wild potato species Solanum berthaultii. This genetic transformation confirmed that in contrast to the Sb AI suppression in CIS-resistant potatoes, overexpressing Sb AI in CIS-sensitive potatoes resulted in less amylase activity and a lower rate of starch degradation accompanied by a lower reducing sugar ( RS) content in cold-stored tubers. This finding suggested that the Sb AI gene may play crucial roles in potato CIS by modulating the amylase activity. Further investigations indicated that pairwise protein-protein interactions occurred between Sb AI and α-amylase StAmy23, β-amylases St BAM1 and St BAM9. Sb AI could inhibit the activities of both α-amylase and β-amylase in potato tubers primarily by repressing StAmy23 and St BAM1, respectively. These findings provide the first evidence that Sb AI is a key regulator of the amylases that confer starch degradation and RS accumulation in cold-stored potato tubers. [ABSTRACT FROM AUTHOR]

Reducing sugar ( RS) accumulation in cold-stored potato tubers, known as cold-induced sweetening ( CIS), is a crucial factor causing unacceptable colour changes and acrylamide formation of fried products. The activity of vacuolar invertase (Stvac INV1) is proved important for the CIS process, and invertase inhibitors are speculated to play roles in the post-translational regulation of Stvac INV1 activity. In our previous research, two putative inhibitors (StInvInh2A and StInvInh2B) of Stvac INV1 were implied to be involved in potato CIS. Here, we further reported that StInvInh2A and StInvInh2B had similar function that specifically inhibited Stvac INV1 activity in potatoes. The genetic transformation of these inhibitor genes in potatoes by overexpression in CIS-sensitive and RNAi-silenced in CIS-resistant genotypes showed that Stvac INV1 activity was strongly regulated by alteration of the transcripts of the inhibitors without impacting on the expression of Stvac INV1. A negative power relationship was found between the transcripts of the inhibitors and Stvac INV1 activity, suggesting 1) a transcriptional determination of the inhibitory capacity of StInvInh2A and StInvInh2B and 2) a significant inhibitory role of these inhibitors in post-translational modulation of Stvac INV1. The results also demonstrated that depression of Stvac INV1 activity through overexpression of StInvInh2A and StInvInh2B weakened accumulation of RS and acrylamide in cold-stored tubers and consequently improved the chip quality. The present research strongly suggest that both StInvInh2A and StInvInh2B function as inhibitors of Stvac INV1 and play similar roles in regulating potato CIS by capping Stvac INV1 activity. These inhibitors could be novel genetic resources applicable for improving quality of potato processing products. [ABSTRACT FROM AUTHOR]

Abstract: The modulation of the activity of enzymes associated with carbohydrate metabolism is important for potato cold-induced sweetening (CIS). A novel RING finger gene SbRFP1 was cloned and its expression was found to be cold-inducible in potato tubers of the CIS-resistant genotypes. Transformation of SbRFP1 in potatoes confirmed its role in inhibiting β-amylase and invertase activity, which consequently slowed down starch and sucrose degradation and the accumulation of reducing sugars in cold stored tubers. These findings strongly suggest that SbRFP1 may function as a negative regulator of BAM1 and StvacINV1 to decelerate the accumulation of reducing sugars in the process of potato CIS. [Copyright &y& Elsevier]

Abstract Potato cold-induced sweetening (CIS) is a major drawback restricting potato process industry. Starch degradation and sucrose decomposition are considered to be the key pathways in potato CIS. Our previous study showed that the RING finger gene SbRFP1 could slow down starch degradation and the accumulation of reducing sugars (RS) through inhibiting amylase and invertase activity in cold-stored tubers. However, the regulation mechanism of SbRFP1 is not clear. In this paper, we first proved that SbRFP1 could promote starch synthesis and modify the shape of starch granules. By further yeast two hybrid, GST-pull down and inhibition of enzyme activity assays, we confirmed that SbRFP1 could slow down the transformation of starch to RS in tubers mainly through the inhibition of β-amylase StBAM1 activity. SbRFP1 was also proved to possess E3 ubiquitin ligase activity by ubiquitination assay. Thus, SbRFP1 may regulate the accumulation of RS in cold-stored tubers by ubiquitination and degradation of StBAM1. Therefore, our study reveals the regulatory mechanism of SbRFP1 in the process of CIS and provides more powerful evidence for the effect of starch degradation on potato CIS. Highlights • SbRFP1 interact and inhibit the activity of StBAM1. • SbRFP1 possess E3 ubiquitin ligase activity. • SbRFP1 regulate the accumulation of RS in cold-stored tubers by ubiquitination and degradation of StBAM1. [ABSTRACT FROM AUTHOR]

A family of NDR1/HIN1-like (NHL) genes that shows homology to the nonrace-specific disease resistance (NDR1) and the tobacco ( Nicotiana tabacum ) harpin-induced (HIN1) genes is reported to be involved in defense. However, little information about NHL genes is available for the potato ( Solanum tuberosum ). Here, we report that the expression of StPOTHR1, a member of the NHL gene family, is associated with resistance in potato against Phytophthora infestans , and is specifically induced in inoculation sites. Overexpression of StPOTHR1 enhances resistance against P. infestans via restricting rapid pathogen proliferation. Further, suppression of StPOTHR1 does not compromise R-mediated cell death. Subcellular localization and posttranscription modifications (PTMs) analysis reveals that StPOTHR1 is localized in plasma membrane (PM) and undergoes multiple PTMs. Moreover, StPOTHR1 interacts with NbMKK5L, a component of the MAP kinase signaling cascade. Taken together, our results suggest that the PM-localized StPOTHR1 contributes to potato immunity against P. infestans and may be associated with the MAP kinase signaling cascade. [ABSTRACT FROM AUTHOR]

Abstract: Dihydrolipoyl acyltransferase (EC 2.3.1.12), a branched-chain α-ketoacid dehydrogenase E2 subunit (BCE2), catalyzes the transfer of the acyl group from the lipoyl moiety to coenzyme A. However, the role of BCE2 responding to biotic stress in plant is not clear. In this study, we cloned and characterized a BCE2 gene from potato, namely StBCE2, which was previously suggested to be involved in Phytophthora infestans–potato interaction. We found that the expression of StBCE2 was strongly induced by both P. infestans isolate HB09-14-2 and salicylic acid. Besides, when the homolog of StBCE2 in Nicotiana benthamiana named NbBCE2 was silenced, plants showed increased susceptibility to P. infestans and reduced accumulation of hydrogen peroxide (H2O2). Furthermore, we found that a marker gene NbrbohB involved in the production of reactive oxygen species, was also suppressed in NbBCE2-silenced plants. However, silencing of NbBCE2 had no significant effect on the hypersensitive responses trigged by INF1, R3a-AVR3aKI pair or Rpi-vnt1.1-AVR-vnt1.1 pair. Our results suggest that BCE2 is associated with the basal resistance to P. infestans by regulating H2O2 production. [Copyright &y& Elsevier]

Highlights: [•] Expression of St-Inh was highest in tubers and was strongly suppressed by cold. [•] Expression of St-Inh in tubers decreased acid invertase activity and RS content. [•] NtInvInh2 has a stronger capacity to inhibit acid invertase activity than St-Inh. [Copyright &y& Elsevier]

Abstract: The activity of vacuolar invertase (VI) is vital to potato cold-induced sweetening (CIS). A post-translational regulation of VI activity has been proposed which involves invertase inhibitor (VIH), but the mechanism for the interaction between VI and VIH has not been fully understood. To identify the potential partners of VI and VIH, two cDNA libraries were respectively constructed from CIS-resistant wild potato species Solanum berthaultii and CIS-sensitive potato cultivar AC035-01 for the yeast two-hybrid analysis. The StvacINV1 (one of the potato VIs) and StInvInh2B (one of the potato VIHs), previously identified to be associated with potato CIS, were used as baits to screen the two libraries. Through positive selection and sequencing, 27 potential target proteins of StvacINV1 and eight of StInvInh2B were clarified. The Kunitz-type protein inhibitors were captured by StvacINV1 in both libraries and the interaction between them was confirmed by bimolecular fluorescence complementation assay in tobacco cells, reinforcing a fundamental interaction between VI and VIH. Notably, a sucrose non-fermenting-1-related protein kinase 1 was captured by both the baits, suggesting that a protein complex could be necessary for fine turning of the invertase activity. The target proteins clarified in present research provide a route to elucidate the mechanism by which the VI activity can be subtly modulated. [Copyright &y& Elsevier]

Abstract: Potato vacuolar acid invertase (StvacINV1) (β-fructofuranosidase; EC 3.2.1.26) has been confirmed to play an important role in cold-induced sweetening of potato tubers. However, the transcriptional regulation mechanisms of StvacINV1 are largely unknown. In this study, the 5′-flanking sequence of StvacINV1 was cloned and the cis-acting elements were predicted. Histochemical assay showed that the StvacINV1 promoter governed β-glucuronidase (GUS) expression in potato leaves, stems, roots and tubers. Quantitative analysis of GUS expression suggested that the activity of StvacINV1 promoter was suppressed by sucrose, glucose, fructose, and cold, while enhanced by indole-3-acetic acid (IAA), and gibberellic acid (GA3). Further deletion analysis clarified that the promoter regions from −118 to −551, −551 to −1021, and −1021 to −1521 were required for responding to sucrose/glucose, GA3, and IAA, respectively. These findings provide essential information regarding transcriptional regulation mechanisms of StvacINV1. [Copyright &y& Elsevier]

Abstract: Really interesting new gene (RING) finger proteins function as ubiquitin ligase and play key roles in biotic and abiotic stresses. A new RING-H2 finger protein gene, StRFP1, was cloned from Phytophthora infestans-inoculated leaves of potato (Solanum tuberosum) clone 386209.10, which is free of R1-R11 genes. The deduced amino acid sequence was characterized by an N-terminal transmembrane domain, a GLD region and a RING-H2 finger signature. StRFP1 is homologous to the tobacco NtACRE132 protein and belongs to the ATL family. The DNA gel blot analysis and mapping revealed that StRFP1, an intron-free gene, had one to two copies in the potato genome and was located on chromosome 3. RT-PCR assays showed that StRFP1 was constitutively expressed in potato plants and significantly induced in detached potato leaves by P. infestans and plant defense-related signal molecules, abscisic acid, salicylic acid and methyl jasmonate. Transient expression studies revealed that StRFP1 fused with GFP localized to the plasma membrane or out of that in onion epidermal cells. The function of StRFP1 in potato resistance against late blight was further investigated by constructing overexpression and RNA interference (RNAi) vectors, which were introduced into potato cv. E-potato 3, respectively. By challenging the detached leaves with mixture races of P. infestans, all of the StRFP1-overexpressing plants displayed slower disease development than non-transformed controls in terms of the lesion growth rate (LGR). In contrast, StRFP1-silencing plants through RNAi were more susceptible to pathogen infection. The present results demonstrate that StRFP1 contributes to broad-spectrum resistance against P. infestans in potato. [Copyright &y& Elsevier]

Abstract: StPUB17, a novel UND/PUB/ARM repeat type gene, was isolated from leaves of potato (Solanum tuberosum L.) clone 386209.10 using the rapid amplification of cDNA ends strategy with the primers designed according to a potato EST fragment up-regulated by Phytophthora infestans. StPUB17 was confirmed intron-free by comparison of the cDNA and genomic DNAs. The RT-PCR analysis showed that StPUB17 was constitutively and differentially expressed in all tissues, and was significantly induced in detached leaves subjected to P. infestans, signal molecules such as salicylic acid, methyl jasmonate, ethylene, abscisic acid and wounding. The gene expression was also strongly up-regulated when in vitro plantlets were exposed to high (40°C) and low (4°C) temperatures and dehydration induced by polyethylene glycol and NaCl. The function of StPUB17 was further clarified by silencing it in potato using RNAi-based post-transcriptional gene silencing (PTGS). The results demonstrated that StPUB17-silenced plants exhibited more susceptible to the infection of P. infestans and more sensitive to the stress of NaCl. Present data indicated that StPUB17 is a gene harboring broad-spectrum responses to both biotic and abiotic stresses in the potato and may play crucial roles in late blight resistance and salt tolerance of the crop. [Copyright &y& Elsevier]

Abstract: To elucidate the molecular events of potato quantitative resistance to Phytophthora infestans, we performed a comprehensive transcriptional analysis using cDNA microarrays, containing 1009 ESTs from a subtractive library. Leaves of a moderately resistant potato clone were inoculated with P. infestans and sampled at nine time points ranging from 2 to 72h after inoculation. A total of 348 P. infestans-responsive genes were identified. These functional genes are mostly related to metabolism, plant defense, signaling and transcription regulation, involving the whole process of plant defense response to pathogens. Based on the general expression patterns of these genes at different time points, we discriminated distinct stages of potato defense against P. infestans and revealed genes participating in each stage. To further understand the dynamics of P. infestans-induced gene expression, hierarchical clustering was used to illustrate their various expression profiles during the time course, including early, mid and late gene induction as well as early gene repression. Interestingly, some genes involved in the hypersensitive response were also identified, suggesting that a same or similar defense system may exist in both race-specific and race-nonspecific resistances. In addition, 114 novel genes with unknown functions were isolated. [Copyright &y& Elsevier]

• The cold responsive gene expression profiles were compared in two potato genotypes. • The AGPase and INVINH expression are important for enhancing CIS resistance. • The candidate transcription factors regulating CIS resistance were identified. Potato accumulates large amounts of soluble sugar during cold storage periods. However, a system based understanding of this process is still largely unknown. Here, we compared the dynamic cold-responded transcriptome of genotypes between cold-induced sweetening resistant (CIS-R) and cold-induced sweetening sensitive (CIS-S) in tubers. Comparative transcriptome revealed that activating the pathways of starch degradation, sucrose synthesis and hydrolysis could be common strategies in response to cold in both genotypes. Moreover, the variation in sugar accumulation between genotypes may be due to genetic differences in cold response, which could be mainly explained: CIS-R genotype was active in starch synthesis and attenuated in sucrose hydrolysis by promoting the coordinate expression of a series of genes involved in starch-sugar interconversion. Additionally, transcription factors, the candidate master regulators of starch-sugar interconversion, were discussed. Taken together, this work has provided an avenue for studying the mechanism involved in the regulation of the CIS resistance. [ABSTRACT FROM AUTHOR]