Evaluation of resistance sources and inheritance of resistance in kidney bean to Indian virulences of Colletotrichum lindemuthianum

Summary Forty nine common bean lines comprising of exotic accessions and locally grown cultivars evaluated against Colletotrichum lindemuthianum exhibited differential resistance to its races in Himachal Pradesh, a north-western Himalayan state of India. Some exotic accessions like G 2333, Cornell 49242, PI 207262, Mexique 222, TO, Perry Marrow, Kaboon and Widusa were resistant to more than five Indian races, whereas two Indian accessions KRC-5 and Hans showed resistance to six and four races, respectively. However, nine accessions KRC-8, KR-40, KR-43, KR-81, KR-62-2, KR-90, KR-142, KR-148, and KR-216 were resistant to three races. Race specific resistance has been observed in different bean cultivars. Studies on inheritance of resistance in exotic accession G 2333 and Indian accession, KRC-5 showed that two independent dominant genes conferred resistance in G 2333 to race 3 and 515 and a single dominant gene controlled resistance in KRC-5 to race 775, indicating resistance from these sources is easily transferable to the locally adapted susceptible cultivars.     

Development of common bean (Phaseolus vulgaris L.) lines resistant to the bean pod weevil,Apion godmani Wagner,in Central America

Summary The larva of the bean pod weevil (BPW), Apion godmani Wagner (Coleoptera: Curculionidae), causes serious yield losses in common bean (Phaseolus vulgaris L.) in Mexico and Central America, by consuming the seed as it develops in the immature pod. Resistance to the BPW was identified in bean germplasm of highland Mexican origin, and these sources of resistance were incorporated into a pedigree breeding program to recover locally adapted lines resistant to Bean Common Mosaic Virus and BPW, with commercial grain for Guatemala, Honduras and El Salvador. These lines yielded as well as or better than local cultivars in the absence of the insect, and better than local cultivars when the BPW was present. Resistance appeared to be governed by several genes, and was stable across geographic areas, seasons and planting systems.     

Genetic and chemical analyses of six cowpea and two <Emphasis Type="Italic">Phaseolus</Emphasis> bean species differing in resistance to weevil pest

The objective of this work was to evaluate the genetic variability among six cowpea (Vigna unguiculata) cultivars differing in their resistance to Callosobruchus maculatus (F.) weevil. Two resistant bean cultivars were used to compare between the sensitive, moderate tolerant, and high tolerant cowpea cultivars. The differentiations were performed by using random amplified polymorphic DNA (RAPD) fingerprinting, protein concentration and organic and non-organic components in seed coat. Six polymorphic primers were identified, resulting in different informative bands. Based on polymorphic profiles, three clusters were formed. Clustering was mainly affected by the resistance to weevil pest. The sensitive cowpea cultivars were separated in one group, the moderate tolerant and high tolerant cultivars came in separate groups, and finally, the resistant bean cultivars separated clearly in one distinct group. The most interesting result was represented by concentration of total protein in the seed coat. The protein concentration in the resistant bean cultivars were approximately 50% less than concentration in each of the moderate tolerant and sensitive cultivars of cowpea. Ferric ions were about 25% less than the moderate tolerant and sensitive cultivars. The concentrations of calcium and potassium in seed coats were higher in the resistant beans than in cowpea cultivars. Cobalt was about four times higher in resistant bean than in the sensitive and moderate tolerant cowpea cultivars, which may play a major role in seed resistance to weevil.     

The kappa race of Colletotrichum lindemuthianum and sources of resistance to anthracnose in Phaseolus beans

Summary The utilization of American and European bean cultivars as host differentials for distinction of races of Colletotrichum lindemuthianum has been discussed. The new race occurring at Ebnet. Germany, since 1973 is named kappa. It broke down resistance derived from the Are gene originating from Cornell 49–242. Resistance to this kappa race appeared to be present in some European and Asiatic bean cultivars as well as in some American bean accessions.     

New sources of resistance of cowpea (Vigna unguiculata) toStriga gesnerioides,a parasitic angiosperm

Summary Thirty-seven accessions of cowpea and yard-long bean were assessed for resistance toStriga gesnerioides. Cowpea plants were grown using anin vitro method, then inoculated with young seedlings ofS. gesnerioides produced from seed from three West African countries. Resistance was assessed by comparing the number and size ofS. gesnerioides tubercles on these accessions with those on a known susceptible cowpea, cv. Blackeye. Two cowpea landraces, APL-1 and 87-2, were completely resistant toS. gesnerioides from Burkina Faso, Mali and Cameroon and partially resistant toS. gesnerioides from Niger. Complete resistance was expressed either as a hypersensitive response of infected root tissues or as a severely retarded development of successful infections. All other accessions, including three samples of yard-long bean were susceptible toS. gesnerioides. The original 87-2 plants segregated for resistance and susceptibility. However, uniformly resistant progeny were obtained by producing seed from vegetatively propagated clones of single resistant 87-2 plants. Resistance of APL-1 and 87-2 toS. gesnerioides was confirmed in pot and field trials. Neither of these cowpeas were resistant toAlectra vogelii. Varieties APL-1 and 87-2 provide additional sources of resistance to most races ofS. gesnerioides, including a newly discovered virulent race from Benin.Abbreviations ICRISAT International Crops Research Institute for the Semi-Arid Tropics - IITA International Institute of Tropical Agriculture - SAFGRAD Semi-Arid Food Grain Research and Development     

Breeding alfalfa cultivars resistant to the alfalfa weevil

A ten-year study consisting of nine field experiments was conducted to evaluate alfalfa (Medicago sativa L.) for resistance to larval feeding by the alfalfa weevil (Hypera postica Gyllenhal). The results were consistent over years and experiments. Only one source of alfalfa germplasm, designated Starnes, showed resistance to larval feeding in the field under natural infestations of the weevil. The cultivars Team, Arc, and Liberty have been developed from this germplasm source. The only effective method of selection for field resistance to the weevil was field selection under natural weevil infestations. Selection was based on low larval feeding on spaced plants. After six generations of such selection the Starnes germplasm showed only one-third as much defoliation as did the check cultivars. The heritability of the weevil resistance was moderately high. Field resistance was transferred to other strains through hybridization; hybrids were intermediate to the resistant and susceptible parents. Resistance was stable.Laboratory and greenhouse efforts to breed alfalfa weevil resistant alfalfa-using such techniques as adult cotyledon-nonpreference tests, adult leaf-feeding tests, larval-growth and survival tests, oviposition-stimulus tests, and oviposition-nonpreference tests-have failed to produce field resistance.Paper No. 5308 of the Journal series of North Carolina Agricultural Experiment Station, Raleigh, N. C., in cooperation wich ARS, USDA. Received 15 June 1977.     

Linkage of dominant hypersensitive resistance to bean common mosaic virus to seed color in Phaseolus vulgaris L.

Summary Evaluation of Phaseolus vulgaris germplasm bank materials and progenies from a large number of crosses using red- or yellow-colored, BCMV-susceptible bean lines, crossed to purple- or grey/brown-colored, hypersensitive-resistant lines, suggested strong trait association between seed color and BCMV resistance. The cross of red-mottled I⁺I⁺ (susceptible) BAT 1255R to isogenic purple-mottled II (resistant) BAT 1255M was made to study the segregation of the two characters and to recover red-mottled resistant progenies. No recombinant genotypes were observed among 353 F₃ families inoculated with BCMV-NL3, suggesting that linkage of purple-mottled seed color and dominant BCMV resistance is very close.Contribution of the Centro Internacional de Agricultural Tropical.     

Field screening ofPisum accessions to evaluate their susceptibility to the pea weevil (Coleoptera: Bruchidae)

Summary Seventeen unreplicated field trials over nine sites and four years were used to classifyPisum germplasm (P. sativum L. &P. fulvum Sibth. & Sm) as potential sources of resistance to the pea weevil,Bruchus pisorum (L.). The emergence of adult weevils from <10% of harvested seed was used as the selection criterion to indicate possible resistance. A total of 1900Pisum accessions were assessed using the field trials and 1754 of theP. sativum accessions were eliminated. However in the 18P. fulvum accessions screened, the level of infestation by pea weevil was always below the arbitrary resistance threshold selected. This suggests thatP. fulvum accessions could be a valuable source of resistance to the pea weevil.     

Breeding the common bean (Phaseolus vulgaris L.) for resistance to bean common mosaic virus: alternatives to backcrossing

A series of field experiments was undertaken in order to determine whether resistance to bean common mosaic virus (BCMV) could be incorporated into genotypes of the common bean (Phaseolus vulgaris L.) suitable for cultivation in Zimbabwe without recourse to backcrossing. Six inbred genotypes carrying the resistance-conferring alleles at the loci I and Bc-3 were crossed with five locally-adapted inbred genotypes. The first experiment comprised F₃ progeny rows, each derived from a single unselected F₂ plant, the second, F₃ bulks selected for resistance, and the third, a comparison of selected and unselected F₂-derived F₄ lines. The number of days to flowering and to maturity, the incidence of mosaic and necrosis symptoms, seed yield and seed size were recorded. There was evidence that late flowering and maturity were associated with BCMV resistance in some crosses, though not strongly enough to present an obstacle to plant breeding. The incidence of virus symptoms and seed yield were influenced by genetic factors additional to the major resistance genes, and variation in seed yield was present not only between bulk populations of crosses, but also between single-row plots of lines within crosses. This indicates that early-generation selection for yield in the presence of BCMV, even among progeny selected for BCMV-resistace, is likely to be effective. However, the variation in yield among F₄ lines was least in the highest-yielding crosses, which may represent a limit to successful selection for yield. Seed size was partly under additive genetic control, but there was also evidence of non-allelic interactions. There was no association between large seed size, preferred by consumers, and susceptibility to BCMV in the progeny, indicating that the association between these characters in the parent lines is fortuitous and will not present an obstacle to plant breeding. It is noted that a considerable amount of useful genetic information can be obtained without recourse to elaborate crossing schemes, provided that unselected progeny are included in experiments as controls. The evidence presented indicates that resistance to BCMV can be combined with appropriate values of maturity date, yield and seed size without the need for backcrossing.     

Introgressing resistance to bacterial and viral diseases from the middle American to Andean common bean

The genetic base of cultivars within market classes of common bean (Phaseolus vulgaris L.) is narrow. Moreover, small- and medium-seeded Middle American cultivars often possess higher yield and resistance to abiotic and biotic stresses than their large-seeded Andean counterparts. Thus, for broadening the genetic base and breeding for higher yielding multiple stress resistant Andean cultivars use of inter-gene pool populations is essential. Our objective was to determine the feasibility of introgressing resistance to Been common mosaic virus (BCMV, a potyvirus), and the common [caused by Xanthomonas campestris pv. phaseoli (Xcp) and X. campestris pv. phaseoli var. fuscans (Xcpf)] and halo [caused by Pseudomonas syringae pv. phaseolicola (Psp)] bacterial blights from the Middle American to Andean bean, using gamete selection. Also, we investigated the relative importance of the use of a landrace cultivar versus elite breeding line as the last parent making maximum genetic contribution in multiple-parent inter-gene pool crosses for breeding for resistance to diseases. Two multiple-parent crosses, namely ZARA I = Wilkinson 2 /// ‘ICA Tundama’ / ‘Edmund’ // VAX 3 / PVA 773 and ZARA II = ‘Moradillo’ /// ICA Tundama / Edmund // VAX 3 / PVA 773 were made. From the F₁ to F₅ single plant selection was practiced for resistance to the common and halo bacterial blights in both populations at Valladolid, Spain. The parents and F₅-derived F₆ breeding lines were evaluated separately for BCMV, and common and halo bacterial blights in the greenhouse at Filer and Kimberly, Idaho in 2001. They were also evaluated for the two bacterial blights, growth habit, seed color and 100-seed weight at Valladolid in 2002. All 20 F₁ plants of ZARA I were resistant or intermediate to common and halo bacterial blights in the greenhouse, but their F₂ and subsequent families segregated for both bacterial blights. Segregation for resistant, intermediate, and susceptible plants for common bacterial blight occurred in the F₁ of ZARA II. Simple correlation coefficient for common bacterial blight between the F₁ and F₁-derived F₂ families was positive (r = 0.54 P < 0.05) for ZARA II. From the F₂ to F₅ the number of families resistant to both bacterial blights decreased in both populations. Only four of 20 F₁ plants in ZARA I resulted in seven F₆ breeding lines, and only one of 32 F₁ plants in ZARA II resulted in one F₆ breeding line resistant to the three diseases. None of the selected breeding lines had seed size as large as the largest Andean parent. The use of elite breeding line or cultivar as the last parent making maximum genetic contribution to the multiple-parent inter-gene pool crosses, relatively large population size in the F₁, and simultaneous selection for plant type, seed traits as well as resistance to diseases would be crucial for introgression and pyramiding of favorable alleles and quantitative trait loci (QTL) of interest between the Andean and Middle American beans.     

DNA markers linked to Pga1, an adzuki bean gene that confers resistance to Cadophora gregata race 1

Brown stem rot (BSR) caused by Cadophora gregata f. sp. adzukicola (syn. Phialophora gregata) is a serious soilborne disease of adzuki bean (Vigna angularis) in Japan. Cultivation of resistant cultivars is the most effective disease control method, therefore the selection of resistant lines is a priority for breeders. BSR-resistant adzuki bean lines have been screened in pathogen-infected fields. However, field selection using the pathogen and artificial inoculation methods is time-consuming and labor-intensive. In the present study, we used 105 F₃ lines derived from a cross between a BSR-resistant cultivar ‘Syumari’ and a susceptible cultivar ‘Buchishoryukei-1’ for BSR inoculation tests. Amplified fragment-length polymorphism (AFLP) analyses with 1024 primer sets revealed that six fragments were polymorphic between resistance and susceptible bulked groups. Five DNA markers (Pg77, Pg118, Pg138, Pg139 and Pg126) were developed from the nucleotide sequences of polymorphic AFLP markers and their flanking regions. Pg118, which was derived from E-ACT/M-ACT-118, was tightly linked to the resistance gene Pga1 and was converted into a codominant marker for its easier use in marker-assisted selection for adzuki bean BSR resistance. Finally, the applicability of the developed markers for BSR resistance was tested on 32 adzuki bean accessions or cultivars.     

A major QTL for common bacterial blight resistance derives from the common bean great northern landrace cultivar Montana No.5

Knowledge of the evolutionary origin and sources of pest resistance genes will facilitate gene deployment and development of crop cultivars with durable resistance. Our objective was to determine the source of common bacterial blight (CBB) resistance in the common bean Great Northern Nebraska #1 (GN#1) and GN#1 Selection 27 (GN#1 Sel 27). Several great northern cultivars including GN#1, GN#1 Sel 27, and Montana No.5 (the female parent of the common x tepary bean interspecific population from which GN #1 and GN # 1 Sel 27 were derived) and known susceptible checks were evaluated for CBB reaction in field and greenhouse environments. These genotypes and CBB resistant and susceptible tepary bean including Tepary #4, the male parent and presumed contributor of CBB resistance toGN#1 and GN#1 Sel 27, were assayed for presence or absence of three SCAR markers tightly linked with independent QTLs conditioning CBB resistance. The parents and F₂ of Montana No. 5/GN #1 Sel 27 and Montana No.5/Othello(CBB susceptible) were screened for CBB reaction and SCAR markers. CBB resistance in Montana No.5 was comparable to that of GN#1 and GN#1 Sel27. The SAP6 SCAR marker present in GN#1 and GN#1 Sel 27 was also present in Montana No.5, and it co-segregated (R ² =35%) with the CBB resistance in the Montana No.5/Othello F₂ population. Although a few CBB resistant and susceptible transgressive segregants were found in the F₂ of MontanaNo.5/GN #1 Sel 27 and later confirmed by F₃ progeny tests, SAP6 SCAR marker was present in all progenies. None of the tepary bean specific CBB resistance-linked SCAR markers were present in GN#1, GN#1 Sel 27, or Montana No.5. A cluster analysis of 169 polymorphic PCR-based markers across three common bean and Tepary #4 indicated that GN#1, GN#1 Sel 27, and Montana No.5 were closely related, and not related at all with Tepary #4.Thus, these results clearly indicate Montana No.5, not Tepary #4, as the source of CBB resistance in GN#1 and GN#1 Sel 27. This revised version was published online in August 2006 with corrections to the Cover Date.     

Screening wild Lycopersicon species for resistance to powdery mildew (Oidium lycoperiscum)

Since the late 1980s powdery mildew, designated Oidium lycopersicum, frequently invaded the tomato crop in Western Europe. All commercial cultivars are susceptible. To screen for resistance in wild species a reliable and efficient disease test was developed. Young plants with two to three true leaves are inoculated at high relative humidity by spraying with a freshly prepared suspension of 2×10⁴ conidia, ml^–1. Symptoms are periodically evaluated according to a scale based on the percentage of leaf area with mycelium.One hundred and twenty seven accessions, representing eight wild Lycopersicon species, were screened for resistance to O. lycopersicum. A large variation in resistance was found between species. L. hirsutum was the most resistant species; L. pennellii was moderately resistant; species of the subgeneric group of L. esculentum and of the peruvianum-complex were all susceptible. L. parviflorum was classified separately due to a large variation between accessions. Except for this species, a low variation was found between accessions within species. High levels of resistance were observed in four accessions of L. hirsutum, in one of L. parviflorum and in one of L. peruvianum. This resistance is characterized by a very low disease incidence and a strongly restricted mycelium growth and lack of sporulation.     

Components of partial resistance in phaseolus beans against an Ethiopian isolate of bean rust

Summary Banana weevil (Cosmopolites sordidus Germar) is a major limiting factor in the cultivation of plantains and bananas (Musa spp. L.) in sub-Saharan Africa. The larvae damage the crop by tunneling in the corm. Chemical control is feasible but not sustainable, whereas host plant resistance is safe and has long term benefits. Banana weevil damage and infestation levels and corm hardness were assessed in Musa germplasm in order to determine the genetic control and potential mechanisms of resistance to this insect pest. Susceptibility/resistance to weevil was measured by the percentage coefficient of infestation (PCI) and damage in cross sections (CS) of the corm. All plantains were equally susceptible to the banana weevil, while a wild banana accession and some cooking and dessert banana cultivars showed high levels of resistance. Differential genotypic responses were observed in euploid plantain-banana hybrids. Segregation results suggest that host plant response to weevil in Musa is controlled by gene(s) exhibiting partial dominance towards the resistant parent and modifier genes with additive and dosage effects for susceptibility in the plantain parent. In natural banana germplasm, resistant clones showed increased corm hardness, as measured by a penetrometer in longitudinal and cross sections of outer and central corm tissues. This might suggest a non-preference mechanism for weevil resistance. However, the lack of correlation between corm hardness with PCI and CS scores in the segregating progenies suggested that other mechanisms may be more important in conferring resistance to banana weevil.     

New sources of resistance to anthracnose and angular leaf spot of beans (Phaseolus vulgaris L.)

Summary over 13000 CIAT bean accessions were evaluated for their reactions to the anthracnose (Colletotrichum lindemuthianum) and angular leaf spot (Isariopsis griseola) pathogens over a 3 yr period. Among these accessions, 156 were resistant to all races of the anthracnose pathogen collected from Popayán, Colombia. Thirty were resistant to numerous races obtained from other parts of the world, including Europe. Although many of these new resistant sources originated in Mexico and Central America, they are quite diverse for geographic origin, plant type, seed color and seed size. In addition, more than 50 of the 156 lines were also resistant to isolates of I. griseola with diverse sources of origin throughout Colombia.     

Genetics of resistance to 3 isolates of Ascochyta fabae on Faba bean (Vicia faba L.) in controlled conditions

Summary Genetics of resistance to Ascochyta fabae Speg. in Vicia faba L. was studied with a final objective to develop resistant faba bean varieties to Ascochyta blight. The study was conducted separately on 3 single spore isolates (AF10-2 and AF13-2 from Tunisia and AF4-3 from France) and belonging to different groups of virulence (GV1 and GV2). Important general combining ability (GCA) effects were found especially with isolates AF10-2 and AF4-3. Specific combining ability (SCA), although significant for the 3 isolates, was important only with AF13 -2, but less important than GCA. Additive gene effects were predominant to non-additive effects. Lines 29H and A8817 transmitted to their progenies resistance to the 3 isolates, whereas 14–12 and 19TB conferred resistance to their progenies only with isolates AF13-2 and AF4-3, respectively. In the material studied, resistance was generally controlled by dominant genes but also could be attributed to recessive genes although less frequent. Analysis of segregation in the F2 of 2 crosses between the resistant lines (A8817 and 29H) and the susceptible line (14–12) with isolate AF4-3 revealed dominant monogenic control at the level of leaves in the 2 resistant lines and, in addition, a recessive gene controlling resistance of stems. Non-allelic interactions were occasionally manifested and their origin appeared to be due to line 19TB. A recurrent selection scheme was proposed with the objective to develop improved open-pollination populations and synthetic varieties responding to the objective of the national Tunisian research programme on faba bean.     

Reaction of Lycopersicon cultivars and wild accessions to tomato leaf curl virus

Summary 1201 tomato cultivars, breeding lines and accessions of Lycopersicon species were screened for tomato leaf curl virus (TLCV) under field and laboratory conditions during summer seasons of 1986 to 1989. Two lines of L. hirsutum (PI 390658 and PI 390659) and 2 lines of L. peruvianum (PI 127830 and PI 127831) were resistant to TLCV infection. These accessions did not produce any leaf curl symptoms either in field or after inoculation by whitefly Bemisia tabaci with TLCV. Adult whiteflies died within 3 days after releasing on resistant accessions (PI 390658, PI 390659, PI 127830), whereas the whiteflies survived upto 25 days on susceptible tomato cultivars. Under field conditions 0–4 and 5–25 adult whiteflies were observed on resistant and susceptible cultivars respectively. Hybridization was effected using the popular tomato cultivars Arka Sourabh, and Arka Vikas, as the female parents and the resistant Lycopersicon wild species as the pollen parents, to incorporate the resistant gene(s) into the edible tomatoes.     

A rapid procedure for screening the resistance of bean cultivars (Phaseolus vulgaris L.) to Botrytis cinerea and Sclerotinia sclerotiorum

Summary A method to inoculate detached bean leaves with Botrytis cinerea or Sclerotinia sclerotiorum without using a carbon-nitrogen source is described. Leaves were incubated in plastic trays that were covered with transparent plastic bags to maintain air humidity above 95%. Spores of either fungus inoculated in water alone were generally unable to cause infection. Additions of KH₂PO₄ (62.5 mM) or of mixtures of inorganic phosphate and glucose (1.1 or 5.5 mM) to the inoculum stimulated fungal pathogenicity. Also spore concentration (2×10⁵ or 2× 10⁶ spores ml^–1) influenced the ability to produce lesions. At 2× 10⁶ spores ml^–1 both fungi were able to infect primary bean leaves when inoculated in an inorganic phosphate solution. These conditions were used for a preliminary screening for resistance of 15 bean cultivars to both fungi. Only partial resistance to the pathogens was found. Since the procedure is simple and does not require blooming plants, it can be rapidly adopted in screening and breeding programs.Abbreviations Pi inorganic phosphate     

Breeding for resistance to yellow disease of hyacinths. II. Influence of flowering time,leaf characters,stomata and chromosome number on the degree of resistance

Summary Analysis of an incomplete diallel design with 14 hyacinth cultivars and 4 wild accessions of Hyacinthus orientalis showed a significant GCA component for resistance to yellow disease. The late flowering hyacinth cultivars King of the Blues and Marconi and the early flowering Hyacinthus orientalis 70129 were found to be the best combiners for yellow disease resistance. Within a cultivar, the degree of earliness itself was not associated with the degree of resistance. Correlation coefficients of leaf characters and degree of resistance showed that hyacinths with short and/or narrow leaves are generally more resistant than those with long and/or broad leaves. Tetraploid cultivars with few, large stomata tend to be susceptible, diploid ones with many small stomata tend to be the more resistant.     

A rapid screening technique to combine resistance to halo blight and bean common mosaic virus in Phaseolus vulgaris L.

Summary Seven bean lines (Phaseolus vulgaris L.) with differential resistance or susceptibility to race 2 of halo blight (Pseudomonas syringae pv. phaseolicola) and a necrosis-inducing isolate of bean common mosaic virus were inoculated with one or both pathogens in combination, to determine the feasibility of dual screening to identify resistance to both pathogens simultaneously. Dual screening yielded the same results as separate screenings. Neither pathogen affected the disease expression of the other. Simultaneously screening for resistance to both pathogens will shorten the recurrent screening-selection cycle of hybridization programs, and accelerate development of resistant cultivars.Abbreviations BCMV Bean Common Mosaic Virus - cvs Cultivars - HB Halo blight - Inoc. Pt. Inoculation point - NLL Necrotic local lesion - React Reaction - SVN Spreading veinal necrosis, System chloro-Systemic chlorosis - VN Vascular necrosis