Firmness is one of the most important fruit quality traits in strawberries.The postharvest shelf life of this soft fruit is highly limited by the loss of firmness,where cell wall disassembly plays an important role.Pr...Firmness is one of the most important fruit quality traits in strawberries.The postharvest shelf life of this soft fruit is highly limited by the loss of firmness,where cell wall disassembly plays an important role.Previous studies demonstrated that the polygalacturonase FaPG1 has a key role in remodelling pectins during strawberry softening.In this study,FaPG1 knockout strawberry plants have been generated using the CRISPR/Cas9 system delivered via Agrobacterium tumefaciens.Ten independent lines,cv.“Chandler”,were obtained,and all of them were successfully edited as determined by PCR amplification and T7 endonuclease assay.The targeted mutagenesis insertion and deletion rates were analyzed using targeted deep sequencing.The percentage of edited sequences varied from 47%up to almost 100%,being higher than 95%for seven of the selected lines.Phenotypic analyses showed that 7 out of the eight lines analyzed produced fruits significantly firmer than the control,ranging from 33 to 70%increase in firmness.There was a positive relationship between the degree of FaPG1 editing and the rise in fruit firmness.Minor changes were observed in other fruit quality traits,such as colour,soluble solids,titratable acidity or anthocyanin content.Edited fruits showed a reduced softening rate during postharvest,displayed a reduced transpirational water loss,and were less damaged by Botrytis cinerea inoculation.The analysis of four potential off-target sites revealed no mutation events.In conclusion,editing the FaPG1 gene using the CRISPR/Cas9 system is an efficient method for improving strawberry fruit firmness and shelf life.展开更多
Mixed orthogonal arrays of strength two and size smn are constructed by grouping points in the finite projective geometry PG(mn-1, s). PG(mn-1, s) can be partitioned into [(smn-1)/(sn-1)](n-1)-flats such that each (n-...Mixed orthogonal arrays of strength two and size smn are constructed by grouping points in the finite projective geometry PG(mn-1, s). PG(mn-1, s) can be partitioned into [(smn-1)/(sn-1)](n-1)-flats such that each (n-1)-flat is associated with a point in PG(m-1, sn). An orthogonal array Lsmn((sn)(smn-)(sn-1) can be constructed by using (smn-1)/( sn-1) points in PG(m-1, sn). A set of (st-1)/(s-1) points in PG(m-1, sn) is called a (t-1)-flat over GF(s) if it is isomorphic to PG(t-1, s). If there exists a (t-1)-flat over GF(s) in PG(m-1, sn), then we can replace the corresponding [(st-1)/(s-1)] sn-level columns in Lsmn((sn)(smn-)(sn-1) by (smn-1)/( sn-1) st -level columns and obtain a mixed orthogonal array. Many new mixed orthogonal arrays can be obtained by this procedure. In this paper, we study methods for finding disjoint (t-1)-flats over GF(s) in PG(m-1, sn) in order to construct more mixed orthogonal arrays of strength two. In particular, if m and n are relatively prime then we can construct an Lsmn((sm)smn-1/sm-1-i(sn-1)/ (s-1)( sn) i(sm-1)/ s-1) for any 0i(smn-1)(s-1)/( sm-1)( sn-1) New orthogonal arrays of sizes 256, 512, and 1024 are obtained by using PG(7,2), PG(8,2), and PG(9,2) respectively.展开更多
基金supported by the Ministerio de Ciencia,Innovación y Universidades and FEDER EU funds(grant numbers AGL2017-86531-C2-1R and PID2020-118468RB-C21)the University of Malaga(grant number B1-2020_09)+1 种基金CS-R was awarded a PhD Fellowship from the Ministerio de Ciencia,Innovación y Universidades(PRE2018-085509)PhD Program Advanced Biotechnology,University of Málaga.
文摘Firmness is one of the most important fruit quality traits in strawberries.The postharvest shelf life of this soft fruit is highly limited by the loss of firmness,where cell wall disassembly plays an important role.Previous studies demonstrated that the polygalacturonase FaPG1 has a key role in remodelling pectins during strawberry softening.In this study,FaPG1 knockout strawberry plants have been generated using the CRISPR/Cas9 system delivered via Agrobacterium tumefaciens.Ten independent lines,cv.“Chandler”,were obtained,and all of them were successfully edited as determined by PCR amplification and T7 endonuclease assay.The targeted mutagenesis insertion and deletion rates were analyzed using targeted deep sequencing.The percentage of edited sequences varied from 47%up to almost 100%,being higher than 95%for seven of the selected lines.Phenotypic analyses showed that 7 out of the eight lines analyzed produced fruits significantly firmer than the control,ranging from 33 to 70%increase in firmness.There was a positive relationship between the degree of FaPG1 editing and the rise in fruit firmness.Minor changes were observed in other fruit quality traits,such as colour,soluble solids,titratable acidity or anthocyanin content.Edited fruits showed a reduced softening rate during postharvest,displayed a reduced transpirational water loss,and were less damaged by Botrytis cinerea inoculation.The analysis of four potential off-target sites revealed no mutation events.In conclusion,editing the FaPG1 gene using the CRISPR/Cas9 system is an efficient method for improving strawberry fruit firmness and shelf life.
文摘Mixed orthogonal arrays of strength two and size smn are constructed by grouping points in the finite projective geometry PG(mn-1, s). PG(mn-1, s) can be partitioned into [(smn-1)/(sn-1)](n-1)-flats such that each (n-1)-flat is associated with a point in PG(m-1, sn). An orthogonal array Lsmn((sn)(smn-)(sn-1) can be constructed by using (smn-1)/( sn-1) points in PG(m-1, sn). A set of (st-1)/(s-1) points in PG(m-1, sn) is called a (t-1)-flat over GF(s) if it is isomorphic to PG(t-1, s). If there exists a (t-1)-flat over GF(s) in PG(m-1, sn), then we can replace the corresponding [(st-1)/(s-1)] sn-level columns in Lsmn((sn)(smn-)(sn-1) by (smn-1)/( sn-1) st -level columns and obtain a mixed orthogonal array. Many new mixed orthogonal arrays can be obtained by this procedure. In this paper, we study methods for finding disjoint (t-1)-flats over GF(s) in PG(m-1, sn) in order to construct more mixed orthogonal arrays of strength two. In particular, if m and n are relatively prime then we can construct an Lsmn((sm)smn-1/sm-1-i(sn-1)/ (s-1)( sn) i(sm-1)/ s-1) for any 0i(smn-1)(s-1)/( sm-1)( sn-1) New orthogonal arrays of sizes 256, 512, and 1024 are obtained by using PG(7,2), PG(8,2), and PG(9,2) respectively.
